GT CNSH Trong SX Va Doi Song

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C th ni rng, CNSH c bit l cng ngh gene tht l k diu, m ra mt trin vng ln lao gip con ngi c th thc hin c hoi bo to ln trong mt tng lai pht trin. CNSH c Nh nc Vit Nam u tin pht trin nh mt trong 4 ngnh khoa hc cng ngh trng im. CNSH c coi l cng c hin i ha ca sinh hc. V bn cht, CNSH t thn phi l mt ngnh khoa hc cng ngh hon chnh, c tnh c lp v khoa hc v v phm vi ng dng, c sc sng ring v tn ti nh mt lnh vc khoa hc cng ngh hin i cng vi cng ngh thng tin, cng ngh in tang gp phn thc y s pht trin kinh t x hi. p ng c yu cu , CNSH mt mt phi c xy dng nh cc ngnh khoa hc hin i, bn cnh c tnh lin ngnh phi da trn nn tng khoa hc ring vng chc v c th khng trng lp vi cc lnh vc khoa hc cng ngh khc. Tht vy, trong th k XXI, CNSH ngy cng chng t l mt mi nhn ca sinh hc hin i. Trong lch s sinh hc th gii cha bao gi nhn loi t c nhiu thnh tu sinh hc mi v c ngha chin lc nh ngy nay. CNSH c ni dung rt phong ph, a dng, ngy cng c nhng thng tin i mi v cp nht. V vy, nhng ngi vit gio trnh CNG NGH SINH HC TRONG SN XUT V I SNG ny khng sao tha mn c ht nhng tri thc ang i hi ngi c v cng khng sao trnh khi c nhng thiu st. Rt mong c s gp chn thnh ca ng nghip v bn c. Cun sch ny c xut bn vi s ti tr ca Ban iu phi D n Gio dc thuc i hc Hu. Chng ti xin chn thnh cm n Ban iu phi D n Gio dc i hc Hu gip v to mi iu kin thun li cho vic ra i cun sch ny. Cng nhn y, chng ti xin chn thnh cm n TS. Trn Quc Dung cn b trng i hc S phm Hu vit cho chng ti chng 2 ,mc 6: Cng ngh sinh hc trong to ging vt nui cho nng sut cao v mc 7: Vector virus sng trong to vaccine th y ti t hp. Xin chn thnh cm n. Hu, thng 03 nm 2005 Thay mt cc tc gi bin son PGS.TS. Trng Vn Lung i hc Khoa hc-i hc Hu
1 CNSH-M u Lung Trng Vn
M u
1. Th no l cng ngh sinh hc (CNSH)
S bng n ca CNSH. Danh t CNSH xut hin vo na cui ca thp k 50-70 ca th k XX, hin nay c dng kh ph bin. Cho n nay, chng ta khng bit ai l ngi a ra danh t ny v xut x t u? Cng ngh sinh hc c th hiu mt cch n gin l cng ngh s dng cc c th sng sn xut cc sn phm hu ch phc v con ngi. Cng c nhiu ngi a ra nhiu nh ngha, song cha c mt nh ngha no bao trm ht ngha ca n. Lin on chu u v CNSH (European Federation of Biotechnology) nh ngha: CNSH l s ng dng thc tin ca cc c th sinh hc hay thnh phn t bo ca chng to ra nhng sn phm phc v cho sn xut v i sng, iu khin mi trng sng. C ngi li nh ngha: CNSH l k thut cao s dng c th sng hay nhng cht tch t c th y to ra hay sa i mt sinh vt, nht l nng cao cc c tnh c gi tr kinh t ca cc loi ng thc vt hay to ra nhng vi sinh vt c kh nng tc ng n mi trng. Va qua c ngi li cho rng: CNSH c coi l ngnh khoa hc cng ngh ca vic chuyn np gene (DNA) vo t bo hay c th ch nhm khai thc mt cch cng nghip cc sn phm ca gene phc v i sng, pht trin kinh t. Theo nhng nh ngha trn c th hiu CNSH theo hai ngha: Ngha rng: bao gm nhiu dng s dng cc sinh vt vo cc mc ch sn xut nh lm ru, lm men bnh m, fromage (phomat), lm tng, chao,.. Ngha hp: CNSH k thut cao l CNSH phn t c s dng nhng k thut hin i ti t hp DNA, bin np gene qua con ng vector plasmid, c nh enzyme, gn enzyme ln mt c cht no , gi yn s dng nhiu ln. Ty thuc vo vic hiu nh ngha rng hay hp m ngi ta phn ra hai loi: CNSH mi (new biotechnology) v CNSH c in (classical biotechnology). Cng ngh sinh hc c in c th coi l CNSH xut hin trong lch s loi ngi rt sm, c th cch y 5.000-8.000 nm, thm ch 10.000 nm. Trong kinh thnh cng ni n qui trnh lm gim, lm ru nho, lm da, n nay chng ta vn cn s dng qui trnh .
Cng ngh sinh hc mi xut hin khi k thut di truyn ra i. Chng ta s c dp i su vo vn ny trong nhng phn sau.
2. Lch s pht trin CNSH

T sau chin tranh th gii ln th hai, CNSH pht trin nh v bo. Cuc cch mng khoa hc k thut c nhng thay i c bn c lin quan n s pht trin ca vi sinh vt hc, ha sinh hc, l sinh hc, sinh hc phn t, di truyn hc phn t, ha sinh hc hu c. Nhiu m hnh nghin cu gip cho vic nh hng ng n s pht trin ca CNSH c bit l sinh hc phn t. Vo nm 1950-1960, trong nghin cu t c nhiu thnh tu to ln, ni bt nht l vn m di truyn. n nm 1960-1962, chng minh c c ch iu ha hot ng gene v sau (1969), tng hp c gene l mt thnh tu to ln trong sinh vt hc. Sau nm 1972-1975, s ra i ca k thut di truyn, to ra s bng n ca CNSH, c th tin hnh nhng sn xut sinh hc bt u nhng thao tc trong ng nghim (in vitro). K thut di truyn to ra mt cuc cch mng trong sinh hc, ng thi n nh du mt bc pht trin trong sinh hc phn t. Nhng thnh tu ca sinh hc phn t dn n nhng thng nht trong nghin cu sinh hc lm sng t nhng nghin cu c bn v nghin cu ng dng. Trc khi CNSH ra i (t nm 1950-1960) cng c nhng bc pht trin nh sn xut vaccine, khng sinh, acid amin. S pht trin ca CNSH li ko, tp trung ln cc vn sinh hc. Hu nh nhng bc tin ln ca sinh hc hin i li m ra nhng kh nng mi thng l hon ton bt ng i vi CNSH. Trc ht phi ni n cc phng php c hon thin nh cng ngh gene (genetic engineering) nhm cu trc li cc chng vi khun nm men vi cc gene l v vi cc c tnh d kin trc. Tc pht trin CNSH nhanh chng mt cch d thng, thc hin qui m cng ngh rng ln v thc n gia sc, v thc phm v c nhng hormone, peptid, neuropeptid, cc cht cao phn t sinh hc phc tp n cc hp cht v c v hu c tng i n gin. Ngy nay, CNSH l cng c c th p dng cho nhiu ngnh kinh t khc nhau nh nng lm ng nghip, sn xut v ch bin thc phm, chn nui th y, y t v sc khe cng ng, sn xut cc dc cht, sn xut nng lng, chuyn ha ha cht, chuyn ha sn phm ph nng nghip v cng nghip, v.v. Nh phng php ha hc dng polyethylenglycol, phng php vt l xung in ngi ta dung hp protoplast, phng php ngm ht
phn vo dung dch DNA, phng php vi tim gene, phng php dng sng bn gene chuyn gene trc tip vo cc t bo khc nhau thc vt. hoc, ngi ta chuyn gene gin tip c thng qua vic s dng cc vector plasmid hoc to phi soma v.v. C th ni rng, CNSH c bit l cng ngh gene tht l k diu, m ra mt trin vng ln lao gip con ngi c th thc hin c hoi bo to ln trong mt tng lai pht trin vi mt thi gian rt ngn.
3. Ha hn ca CNSH vi cc nc ang pht trin

Trc cuc gp g vi cc em hc sinh trng PTTH, khi cc em hi nh bc hc ni ting, vin s tr tui nht ph ch tch vin Hn lm Khoa hc Lin X (c) Iu. Ovchianhicov: Ti sao vin s li hin dng i mnh cho sinh vt hc? Vin s c ly lm tic v iu khng? Nh bc hc mm ci v ni: * Khng, ti khng tic V sau gii thch: * Vng, chc l c nhng khoa hc khng km phn quan trng hn sinh vt hc. Nhng ti khng bit c khoa hc no khc li quan trng hn sinh vt hoc. Cu tr li hon ton ng n v tt nhin cha ng trong lng say m v tnh yu ca nh bc hc i vi lnh vc hot ng sng to c la chn. Vin s xc nh mt cch su sc v r rng v tr khoa hc v s sng, v tnh qui lut vn ng vt cht sng trong h thng khoa hc c bn, phc tp v hin i. Cch y hn 40 nm, khi tr li phng vn ca nh khoa hc th gii v tng lai ca di truyn mt nh khoa hc v sinh hc phn t ni: Kh m tin on, nhng ch bit n nn 2000 trong mt bui sng ma xun, th sinh ca ti s tr li c cu hi bng cch bin i di truyn th no v chuyn gene ra sao nhng cy u H Lan i chiu cun ngc li t phi sang tri trn gi , sao cho ton b cc l hng c nh sng mt tri to iu kin cho quang hp c tt nht. V cng bng cch chuyn gene nh th no c th bc thuc gene cha cho mt hong t mc bnh tm thn. Ngy nay, cng ngh gene gip cho vic chuyn gene u vit vo vic to ging mi, ghp cc gene tng sc khng ca cy nh to ra nhiu cht c ch s tiu ha ca su b, ngi ta cng chuyn gene protein capsid (nhng khng th ca cy) c th chng c cc virus.
Ngi ta cng dng sng bn gene a nhng gene chng chu iu kin bt li ca ngoi cnh vo c th chng hn hn, chng su bnh, v.v. Bng phng php chuyn gene di truyn, ngi ta bin b sa cho b yaourt, bng phng php dung hp protoplast ngi ta to ra nhng cy va n c (c khoai ty) va n qu (qu c chua), sn xut vaccine ti h hp, lm phng i gene vi k thut PCR (polymerase chaine reaction) thu c nhiu kt qu qu bu. c bit ngy 26/6/2000, cc nh khoa hc thuc d n lp bn gene ngi, mt d n a quc gia do Anh, M ti tr v cng ti Celera Genomics (CG) ca Craig Venter cng cng b bn gene (BG) ngi v c nh gi tng ng vi vic nh du hnh v tr M Neil Amstrong t bc chn u tin ln mt trng vo nm 1969; v hn c thnh tu tm ra thuc khng sinh . l mt thnh tu to ln nht trong lch s di truyn hc, sinh hc phn t v y hc phn t k t khi Watson v Crick cng b cu trc xon kp ca phn t DNA nm 1953. Trc cc nh khoa hc c tnh t nht phi n 2005 mi thit lp c BG cho khong 80% cc gene trong h gene ngi vi kinh ph t nht l ba t USD. Trong thc t, cc nh khoa hc cng b BG ngi vi 97% v n nm 2002 ngi ta gii m hon ton BG ngi. Ngi ta pht hin rng, trong con ngi ch c 30.000 n 35.000 gene (trc y ngi ta cho rng trong con ngi c t 60.000 n 100. 000 gene). Mt s i tng khc li cn cao hn nh la c 50.000 gene. Nhn chung th c n 98% gene tng ng.
Bn gene
Hgene ng i
Tip theo , ngi ta pht hin nhiu gene c kh nng tr nhiu bnh him ngho cho con ngi. Hn th na, ngy 27/12/2002 Gim c iu hnh cng ti sinh sn v tnh (SSVT) Clonaid-Brigitte Boisselier cho bit nhm nh khoa hc thuc cng ti ny ln u tin thc hin thnh cng ca SSVT vo ngy 26 thng 12 nm 2002 v cho ra i b gi t tn l Eve. Khng nh, khng bng hnh, khng tit l danh tnh ngi ph n 31 tui thc hin ca SSVT, Boisselier ni rng clonaid s cung cp chng c bng mu DNA trong 8-9 ngy k t ngy cng b. Xt gc khoa hc, ngi ta cn bn tnh bn nghi thng tin trn. Nhng gc x hi, s in r trong tng c nng ln tm tn gio ca Clonaid th khng ai ng vc.
Vic nhn bn v tnh con Cu Dolly ni ting mt thi (thng 2 nm 1997), nay Cu Dolly cht sau 6 nm tui (cng b ngy 15/2/2003). Phn tch th thy n 12 nm tui v ly t bo t m n c 6 nm tui, sau ny ngi ta cn nhn bn nhiu ng vt khc nh chut, mo, d, ln.Gn y, ngy 7/8/2003, TS Golli ngi Italia thc hin vic nhn bn thm con nga. Vic nhn bn v tnh cc ng vt m ra mt hng mi trong vic bo tn ngun gene qu him ca cc ng vt c nguy c dit chng v ang dit chng. Thi gian gn y ngi ta cng nui cy t bo gc (stem cells) .Khi phi cn giai on rt sm mi c 8 t bo th mt t bo u c kh nng pht trin thnh mt phi hon chnh hoc phn ha thnh bt k loi t bo no ca c th sau ny. Nhng t bo ny c gi l t bo gc nguyn pht. nhau thai mt s t bo cng cn duy tr c kh nng phn ha tim nng v c th nui cy thnh dng t bo gc th pht. Ngi ta cng ng dng cng ngh nano sinh hc (bionanotechnology) cho php thu nhn nhng thng tin v h thng sinh hc mc lng t, u d kch thc nano ti kch thc mt phn t ring r dng trong chn on bnh. Cng ngh nano l phng php in stitu mi cung cp thng tin tt hn v chc nng t bo, l cng ngh thao tc ci bin 2 chiu v 3 chiu i vi m v t bo, vn chuyn v phn phi thuc hoc gene vo m v t bo thng qua khng ch kch thc ht, hot ha v gii phng cht thuc qua c ch v thit b nh bm kch thc nano, van t bo vo c quan nhn to. Vit Nam, chng ta cng dng phng php trc tip bn gene v phng php gin tip chuyn gene bng con ng plasmid a gene chng chu ry nu vo cy la (vin La ng bng sng Cu Long), cy ma chu hn (vin CNSH H Ni) t kt qu bc u. X nghip Dc TW cng chuyn np gene ch vaccine c kt qu. Gn y, ngy 24 thng 2 nm 2004, tip theo vin CNSH H Ni, vin Pasteur thnh ph H Ch Minh cng gii m thnh cng b gene H5N1 (gy bnh cm ngi t g) c hng iu tr bnh ny. Nhng thnh tu khoa hc hin nay, nhng kinh nghim ca th gii chng minh rng: ngi ta ngy nay ch n nhng g xy ra trong ton b sinh hc v nht l trong nhng lnh vc ring ca sinh hc CNSH. ngha xut sc ca CNSH l ch nh sc mnh a dng ca mnh m n m ra nhng con ng mi m gii quyt hng lot
cc vn c tnh ton cu nh tnh hn ch v mi e do thc s ca tiu hao cc ngun nng lng, thc phm truyn thng v cui cng l s nhim mi trng xung quanh. i vi cc nc ang pht trin, CNSH l mt vn then cht, m vn cc nc ny b coi l khc bit so vi cc nc c nn cng nghip pht trin. Thi gian qua, trong cc nc ang pht trin c mt s nc vn ln v t trnh khoa hc cng ngh cao. H c mt nn tng cng ngh vng v mt th trng rng m bo lm ch mt s mi nhn CNSH hng chng vo phc v cc nhu cu ca nc mnh. Tuy nhin, a s cc nc ang pht trin cn ang thiu ngun vn khai thc cc cng ngh , thiu h tng c s cho nhiu nghin cu c bn, ng dng v thiu ngi c trnh cn thit cho cc ngnh cng nghip sinh hc. V vy cc nc ny phi kt hp hi ha nhng tin b ca CNSH vi tnh trng thiu vn nhng li d tha lao ng, nhng b quyt ca CNSH, qui trnh CNSH c truyn v.v. Hin nay cc nc ngho nht v km pht trin v mt cng ngh v khoa hc cng c th thu c mt s li ch do tin b ca CNSH v tham gia vo cuc cch mng CNSH nh cc mng li hp tc quc t v khu vc. Ring khu vc chu , mt s trung tm CNSH ra i nh trung tm T liu Th gii v cc Vi sinh vt MIRCEN Nht Bn, vin CNSH ca i hc Osca, vin Nghin cu Khoa hc K thut Thi Lan (cho vng ng Nam ), trung tm New Delhi nghin cu v c nh N 2 sinh hc, tnh chng chu cy lng thc, ci thin v phn phi cht dinh dng trong thc vt, tng trng v ti sn xut gia sc, pht vaccine phng bnh nhit i. Vin Nghin cu Cao su bng nui cy m Malaysia (RRIM), cng ti Mc in v Ha cht Dainippon (DIC) Tokyo Nht Bn chuyn sn xut cc cht sinh hc tinh khit v cc cht mu thc phm, thc n cho c, m phm t cc loi to. v.v. Cng ngh sinh hc c tm quan trng to ln, v vy, CNSH tr thnh mt trong bn mi nhn ca th gii ngy nay (in t v tin hc, nng lng, vt liu mi, cng ngh sinh hc). Vit Nam, ng v Nh nc ta cng thy r tm quan trng ca CNSH. Vn kin Hi ngh ln th VII ca Ban chp hnh TW ng kha 2 cng nhn mnh: u tin v ng dng pht trin cc cng ngh tin tin nh: cng ngh thng tin phc v yu cu in t ha v tin hc ha nn kinh t quc dn; CNSH trc ht phc v pht trin nng, lm,
ng nghip, ch bin thc phm, dc phm v bo v mi trng sinh thi; cng ngh ch to v gia cng vt liu, nht l ngun nguyn liu trong nc (bi pht biu ca /c nguyn Tng B th Mi ti Hi ngh ln th 7 BCH TW ng kha VII ngy 25/7/1994 trang 84). Trong cc i hi VIII, IX, ng ta cng rt ch trng n vn CNSH. Trong Hi ngh Cng ngh Sinh hc ton quc 2003 (ngy 1617/12/2003), trong nh hng nghin cu v trin khai ca vin CNSH thuc vin Khoa hc v Cng ngh Vit Nam, PGS.TS. Trn L Bnh Vin trng vin CNSH t vn : CNSH c nh nc Vit Nam u tin pht trin nh mt trong 4 ngnh khoa hc cng ngh trng im. CNSH c coi l cng c hin i ha ca sinh hc trong vic phc v pht trin nng lm ng nghip, bo v sc khe cng ng v mi trng bn vng.V bn cht, CNSH t thn phi l mt ngnh khoa hc cng ngh hon chnh, c tnh c lp v khoa hc v v phm vi ng dng, c sc sng ring v tn ti nh mt lnh vc khoa hc cng ngh hin i nh cng ngh thng tin, cng ngh in tang gp phn thc y s pht trin kinh t x hi. p ng c yu cu , CNSH mt mt phi c xy dng nh cc ngnh khoa hc hin i, bn cnh c tnh lin ngnh phi da trn nn tng khoa hc ring vng chc v c th khng trng lp vi cc lnh vc khoa hc cng ngh khc nh cng ngh gene, cng ngh t bo ng thc vt v vi sinh vt, cng ngh enzyme v protein. Mt khc, CNSH phi c mc tiu v ni dung nghin cu c trng ring, l xy dng v pht trin ngnh Cng nghip sinh hc vi chng loi cng ngh v hng ha mang du n c th ca CNSH m nhng nh hng hon thin v chuyn giao cng ngh phc v sn xut. R rng, CNSH l ci cha kha m ng cho s pht trin nn kinh t ca t nc. Cuc cch mng khoa hc k thut ni chung v cuc cch mng CNSH ni ring thu ht nhiu ngi trn tri t ny tham gia vo s nghip cao c . Vin s N.N. Semionov vit rng: c im c bn ca khoa hc th k th XX l ch, n khng cn l ngi n t ca sn xut m tr thnh ngi m ca sn xut. Sinh hc chim mt v tr nh th. Tip sau l vt l hc v ha hc.
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CNSH phc v nng lm ng nhip Lung Truong Vn
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CNG NGH SINH HC PHC V NNG LM NG NGHIP Chng I:
Cng ngh sinh hc vi nng lng

1. T nng lng mt tri n nng lng sinh hc.
Chng ta phi mang n mt tri v tt c s giu c ca th gii hu c quanh ta. Tia sng mt tri tng tc vi cht dip lc ca cy xanh to ra s k diu ca quang hp. T cc cht v c n gin ca t nhin nh nc, CO2 ca khng kh, mui N2, phosphor,thc vt to ra cc cht hu c rt phc tp v cu trc (tc l c trng cho c th sng v tham gia vo thnh phn ca cc c quan v cc m ca chng) l ng, acid amin, nucleotide, vitamin, Nh vy, thc vt hp th nng lng nh sng mt tri, to ra cc cht dinh dng, l hin tng quang hp. Ni mt cch khc, quang hp l mt qu trnh bin quang nng thnh ha nng v nng lng c tch ly li trong cc hp cht hu c. Mt kg cht kh hu c c d tr trong 4.000 kcalo,. Tng lng cht hu c do thc vt tng hp c trn tri t hng nm c 4,5.1011 tn (tnh ra bng ng glucose). Hng nm con ngi ch s dng 3,5% cht hu c do thc vt cn tng hp c, cn cht hu c do thc vt nc tng hp c, con ngi s dng cn ang t. Ngoi ra, trong phn ng quang hp cn gii phng ra O 2 rt cn cho h hp ca mi sinh vt v cho cc qu trnh oxyha khc (hng nm trn tri t cy thi ra trong khng kh 15.104 tn phn t oxygen). Do , tia sng mt tri l c s nng lng ca mi s sng. Mt tri cung cp mt cch rng ri nng lng cho con tu v tr ca chng ta tri t. Theo tnh ton ca cc nh bc hc th hnh tinh ny mi nm nhn c t mt tri khong 5.1019 kcalo. S lng nhit nng ny sn xut ra mt nng lng in khong 2.1026 kw/h, tc l bng s lng
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in to ra trong mt nm ca khong 9 triu nh my in c cng sut tng ng vi nh my thy in Brataki. Tt c th gii cy xanh ca tri t ch s dng ht c mt phn nh do nng lng mt tri a ti: 1-2%. Cc nh bc hc kin tr tm kim cc con ng cho php nng cao hiu sut quang hp d ch thm mt vi ln (thc t mt s nc mt s cy trng cng c h s s dng quang nng trong quang hp l 2-4%), chc hn l iu s mang li nhng li ch khng thua km g vic chim lnh nng lng nhit hch. Cng cn nhn mnh rng, mc d trong tng lai chng ta s khai thc ngun nng lng ht nhn s dng, song mt tri vn l nng lng ch yu i vi s sng trn tri t. ng nh nh vt l hc ngi Php Pierre Curie pht biu (1949) Mc du ti vn tin tng lai ca nng lng nguyn t v thy r tm quan trng ca pht minh ny, tuy nhin, ti cho rng cuc cch mng thc s trong nng lng hc s n ch lc no m chng ta c th thc hin c s tng hp hng lot cc phn t tng t nh dip lc hoc cht lng cn tt hn. Mun t c mc ch , trc ht cn nghin cu t m kiu phn t v tc dng ca quang hp. y chng ta cha k n tr lng thc vt ha thch cng rt ln. Ch mi tnh ring d tr C trong than , du ha v cc kh thp t ti 1018 tn (trung bnh 200 tn/ha v qu t). Theo thng k cha y th d tr C trong cc cht hu c ca sinh vt, trong cc cn b cht hu c ca cc sinh vt cht, trong ha thch do hot ng ca quang hp trc y ca thc vt to ra cng t ti 6.1015 tn.
2. Cc bin php nng cao hiu qu s dng nng lng mt tri v to nng lng bng bin php sinh hc
Hin nay chng ta phi s dng mt cch khn ngoan hn, trit hn cc ca ci m qu trnh quang hp ang to ra hin nay v to ra t hng chc, hng trm, thm ch hng triu nm trc y. Nhng kho bu ny khng m xu. Phn ln chng cha c s dng hoc s dng khng c tt, nu tn dng ht hiu sut quang hp th thc vt bin, i dng, sng ngi, lc a cng c th dng nng lng mt tri tng hp ra mt s lng cht hu c to ln bit bao (A.A. Nhishiporovitch). Ch tnh ring trn cn (khong 1/3 bc x chung ca mt tri chiu xung hnh tinh chng ta) mi nm tng hp c 53 t tn cht hu c, trong trn ng rung: 11 t tn, trn tho nguyn v trn ng c: 6 t tn v trn rng: 36 t tn. Ch mt phn rt nh khi lng vt cht thc vt to ln ny c con ngi dng lm thc n trc tip hoc thc n gin tip (di dng
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cc sn phm c ngun gc ng vt). lm thc n cho con ngi ch dng ht 6% cc sn phm quang hp c to thnh trn ng rung 0,03% sinh khi c to ra bi thc vt trn tho nguyn v trn ng c v ch khong 0,03% sinh khi c to ra trn rng. bin v i dng ni nhn 2/3 bc x mt tri tng hp ra t nht cng khng km phn sinh khi cn, nhng ch mt phn nh dng lm thc n trc tip hoc thng qua tm, c, ng vt m lm thc n cho con ngi. Ngoi vic s dng ci sn c ca vt cht do quang hp to ra, chng ta cn nng cao hiu qu ca b my quang hp nh tm nhng test th nhanh pht hin nhng dng c hiu qu quang hp cao trc ht l dng nhng loi vi To. Hoc, i su vo vic tm hiu c ch di truyn nht l di truyn quang hp b my lc lp hot ng c hiu qu cao hn, bng nhng k thut ti nc, bn phn hp l, chn ging cy trng c nng sut cao, phm cht tt, chng chu gii. p dng cc k thut in vitro nhn nhanh cc ging cy trng. Theo d bo ca mt cng ti t vn khoa hc ging cy trng quc t, sn lng lng thc th gii s tng 5-10% trong vng vi nm ti ch ring nh p dng CNSH (Withen v Anderson, 1986; Faillin, 1986)
Bng I.1. Sn lng hin nay v tng lai ca mt s cy trng nng lm nghip Cy trng Sn lng hin nay Sn lng tng lai (tn/ha) (tn/ha) Ma 70-90 150-200 Sn 15-20 60-100 C chua 20-40 60-100 C du 2-5 10-12 Lc 1,6 4,0 Thu du 0.6 2,5 Thng (n i) 6,8 20-30 Thng (nhit i) 12-20 40-60 Cy l rng (nhit i) 10-20 40-100 Tre 25 100
Ngoi nhng cy lng thc thc phm cung cp nng lng cho con ngi trong ba n hng ngy, chng ta cng cn tn dng mt s cy khc, c ngun nng lng phc v cho i sng x hi, nh nhng cy c du (cy Da, C du, Jojoba) cy c nha m dng lm cht t thay du m, sinh khi cc loi cy ly m ny khong 10 tn/nm tng ng 1,5 tn du m, cy ly tanin (Jojoba). V to ngun nng lng nhng cuc th nghim tin hnh ti Brazil, Trung Quc hay n cng nh cc nc ang pht trin khc
12
cho thy c th kt hp hoc lin kt vic sn xut nng lng vi vic sn xut nng nghip v thc phm ci thin iu kin sinh dng nng thn bng bin php CNSH. Brazil, chng trnh Pro-alcohol pht ng t nm 1975 lm tng trong mt thi k tng i ngn sn lng ethanol ch yu t phng thc cho ln men ng ma. Sn lng t ti 8 t lt ethanol hng nm vo nm 1984 (Larovier, 1985). Mc tiu th trong nm 1985 l 9 t lt v nm 1986 l 12 t lt ethanol (hai triu bn trm nghn t trong s 8.200.000 chic tiu th loi nhin liu cha 20% ethanol). Vic sn xut alcohol nguyn liu ch yu l mt r ng, sn, dch p cy cao lng ngt (Sweet sorghum), c ci ng. Con ng to kh methan (biogas): Trung Quc sn xut kh sinh hc (biogas) bt u trong nhng nm 1950 vi 5 triu b sinh methan c xy dng tnh T Xuyn trong tng s trn 7 triu b khp c nc. Chng trnh biogas lc u ch nhn mnh vo vic thit k v vic ch to b hn l kha cnh vi sinh vt hc (iu kin ln men vi sinh vt hc v nhng vi khun sinh kh methan v khng sinh methan). Nhng nghin cu Thng Hi sa cha nhng khuyt tt ny v tm ra hng s dng nng thn (Chiao, 1986, theo [8]). Hi ngh vi sinh vt biogas t chc nm 1981 v 1983 dnh nhiu hn cho cc mt nghin cu c bn. Ngi ta thy cc vi khun sinh hydrogen v nui cy hn hp lm giu cc vi khun methan sn sinh ra lng methan ln hn nhiu (Sun et al., 1981, theo [8]). Ngi ta cng nhn mnh cc yu t khng sinh methan gi vai tr h trng trong vic sn xut biogas. Hn na, vic tch cc chng tinh khit ca vi khun sinh methan nh: Methanosarcina, Methanobrevibacter arboriphilus, Methanobacterium formicium, Methanococcus mazei lm r hn vic sn sinh biogas v lm tng hiu qu ca qu trnh (Chiao, 1986, theo [8]). Chng trnh biogas Trung Quc chng nhng lm ci tin vic sn xut v tiu dng nng lng cho gia nh m cn nhm kt hp ngy cng mnh vic sn xut lng thc, lp li chu trnh nhng thi b hoa mu v ngn cn vic gy nhim. Cc h thng phi hp sn xut nng lng v thc phm c pht trin cc lng mc. Ti Xin Bu x Lelin, trn ng bng sng Chu Giang gn Qung ng, 1.500 lng ang dng l ch to c bit sy g vi nhit lng 35-40% l un nc bng sc nng mt tri, t trn mi nh cung cp 60-100 lt nc 50 oC hng ngy v ma ng v 70oC v ma h. Nh vy, tit kim 20-30% methan ty mi gia nh s dng. C ni dng biogas lm l sy kh cc tri nui Tm. Ngoi ra, nhng cn b d tha t vic ln men methan, cht thi c dng lm phn bn trng Nm, dng lm thc
13
n cho c, gp phn lm sch sn phm ph v loi b cc ph thi (Larovier, 1985). Ngi ta cng c th s dng To n bo sn xut hydrocarbur nh Botriococcus baurii. To ny c mt s nha khoa hc Php ( trng i hc cao cp Quc gia) nm 1976 quan tm. Trong To c cha lng hydrocarbur t 15-17% trng lng kh. y l loi To nc ngt, nui trng trong iu kin mi trng dung dch tt c th thu c 60 tn cht kh/nm, em n chng ct nh, c th dng nh du m. Cc cht t tch ly phn t bo v, em li tm mnh, cc cht ny s tch ra, t bo vn cn sng c th em nui li. Hin nay ngi ta thy rng, v t bo ny cha nhng cht tng t nh du m nn dng n lm m hnh nghin cu qu trnh to thnh du m. Trn c s nghin cu v c ch ca qu trnh quang hp, c bit l cc cu trc ca lc lp lin quan n chc phn ca n nh th no v vic hp th nng lng nh sng mt tri, bin nng lng thnh dng nng lng ha hc ra sao (phn ny s ni r chuyn Quang hp v nng sut thc vt) ngi ta ch to ra mt s b phn (my mc) s dng nng lng mt tri nh pin sinh hc: mt hp cht cha nhng sinh vt to nng lng (do Potter ch to, 1925). Nhng in cc platinum v dch nui cy ym kh hoc nm men Saccharomyces cerevisiae hoc E. coli t bo xut hin in th m xut hin in cc dng ca platinum. in cc dng platinum ny c t trong mi trng v trng O2. Nu c ln men s to nn dng in c in th 0,30,5 volt, cng dng in 0,02 mA.
Nm 1950-60, cc nghin cu ny c hon chnh, n nay bt Nma vo con tu v u 1950-1960 tr s dng. Sau ny ngi ta s dng nhng cht khng tch in nh glucose: Glucose vi sinh vt ln men H2
4
4H+ OH2H2 2H2O
H2
O2
Anode Cathode Mng Pin sinh hc n gin H2 O2
Ngi ta cng c th dng enzyme bin i mt s cht to H2.
14
H2
Ngoi vic to ra ngun in sinh hc (pin sinh hc ), ngi ta ang nghin cu s dng cc mng quang hp. Nh ta bit, cc sinh vt c kh nng to nng lng. Ngi ta c th c nh t bo thu nng lng. V d; nhng vi khun quang hp nh nh sng to thnh H2 + ATP. To lam Cyanobacteria H2 + NADPH2 m NADPH2 l cht tch tr nng lng (NADPH2 NADP cho ta 4ATP). thc vt thng ng trong lc lp c mng thylakoid l ni to ra H2, H2O2, NADPH2. Cng c nhiu th nghim bin nng lng mt tri thnh in nng. Nht Bn, ngi ta s dng in cc l oxyd titan. TiO2 c bao bi h thng quang ha I (PsI) c chiu sng . Cht kh c dng l ascorbate hay hydroquinon. Phn ng nh sau: P700 + TiO2 P+700 + TiO2 cb (condition barid = dy dn) P+700= + ascorbate P700 + dehydroascorbate P+700 + hydroquinon P700 + quinon Bng cch ny c th to c dng in 100 mA. Dng in to c khng ln nn phi s dng in cc tinh vi - in cc c bc bng protein. P700 kh phc tp nn trong tng lai s c nh mng thylakoid bin quang nng thnh in nng v s s dng trong cc dng c tinh vi nh my in ton. V mt k thut nh hng cho con ngi trong tng lai l nghin cu s dng trc tip bin quang nng thnh in nng.
15
Hin nay, quang nng ha nng (du, than , kh t) nhit nng (t nng) c nng (quay) in nng. Nu tm cch chuyn thng quang nng in nng th s tit kim rt nhiu v trnh vic to nhit nng t nng v s gy nhim. Cn quang hp, thc hin c nh thc vt s lm tinh sch khng kh. Ngy nay, ngi ta cng ang nghin cu ch to cc biosensor, biochip l nhng protein thu nhn nh sng s dng trong my in ton v trong cc dng c tinh vi khc.V d: Anh dng Rhodopsine thu nhn nng lng lng t chuyn vo t bo thn kinh to thnh dng in sinh l. vi khun Halobacterium halobium c cht bacteriorhodopsine nm mng ngoi ca t bo. Bacteriorhodopsine hp th lng t ca nng lng nh sng mt tri to nn nhng bin i lm cho mng c s chnh lch gradien lm bm photon v to nng lng. Ch c loi protein c kh nng thu lng t nh trn th tng lai s c ch to cc b phn ny s dng trong cc my in ton. ng nhin, lm c vic ny s c s phi hp ca cc nh sinh vt, ha hc v in ton. TI LIU THAM KHO 1. Trng Vn Lung, 1999. Chuyn Quang hp v nng sut thc vt. T sch i hc Khoa hc Hu. 2. Trng Vn Lung, V Th Mai Hng, 1999, Gio trnh l thuyt Sinh l hc thc vt. T sch i hc Khoa hc Hu. 3. Nguyn Duy Minh, 1981. Quang hp.Nxb Gio dc H Ni. 4. V Vn V, V Thanh Tm, Hong Minh Tn, 1998. Sinh l hc thc vt. Nxb Gio dc H Ni. 5. Grodzinski A.M., Grodzinski .M., 1964. Sch tra cu tm tt v Sinh l thc vt. Nguyn Ngc Tn v Nguyn nh Huyn dch nm 1981. Nxb Mir Moskva v Nxb Khoa hc v K thut H Ni. 6. Heath O. V. S., 1972. Photosynthes. Izd. Mir, M. 7. Oparin A.I., 1967. C s Sinh l hc thc vt. Tp 1. L c Din v nhng ngi khc dch nm 1975. Nxb Khoa hc v K thut H Ni. 8 Sasson Albert,1988. Biotechnologies and development Cng ngh sinh hc v pht trin. Ngi dch: Nguyn Hu Thc, Nguyn Ln Dng v mt s dch gi khc. Nxb Khoa hc & K thut H Ni.
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16 CNSH phc v nng lm ng nghip Lung Trng Vn
Chng II
Cng ngh sinh hc phc v nng lm ng nghip

1. CNSH c truyn trong vic to ging mi
1.1. Chn lc t nhin T xa xa con ngi cng bit chn ging cy trng, ngay c thi k n lng l, cch y 5000-6000 nm.Theo ti liu ghi chp c th nh chn ging u tin c thc chu u l Lecourteur ngi Php o Gerseille (1 hn o gia Anh v Php) vo u th k th XIX. Mt hm ng mi mt ngi bn Espain (Ty Ban Nha), l mt nh thc vt hc tn l Lagaska i tham quan ng rung la m. ng bn thy la m tuy gieo cng mt ging nhng li c cy rt khc nhau. Theo gi ca bn, Lecourteur em gieo ring 23 dng khc nhau. ng chn c mt ging mi. l phng php chn lc. Bng phng php ny, v sau nhiu nh chn ging chn c nhiu ging mi t vi mun ca con ngi. C s khoa hc ca vic to dng mi ny l qua qu trnh pht trin c th, trong iu kin bt li ca mi trng c th no khng chu ng oc th b tiu dit. Trong iu kin bt li c mt s to ra mt s cht chng chu vi mi trng lm thay i cu trc v hnh thi, ci bin kiu gene v kiu hnh ca qun th theo hng thch nghi v to ra loi mi. Hc thuyt J.B.Lamark (1744-1829) v nht l ca Ch.Darwyn (1809-1882) v ngun gc cc loi l c s bin d c th. Quan im v s sng st ca nhng c th thch ng l ht nhn ca thuyt chn lc t nhin ca Darwyn. 1.2. Lai hu tnh Nm 1694, Kameriarux ngi c pht hin ra cy c cng c ging c ging ci nh ng vt. n nm 1717, nh lm vn ngi Anh l Fershai to c ging hoa Cm chng u tin bng cch lai 2 ging c mu sc khc nhau. Khoa hc v bin d di truyn c Darwyn (1858) v Mendel (1865) pht hin ra nhim sc th, DNA, gene, cc nh khoa hc c mt l lun vng chc v di truyn hc, lm c s cho vic chn ging cy trng. Cho n nay, cc nc tin tin c tp on gingchuyn gi ging cung cp cho cc nh chn ging lm thc liu ban u. Thng mi ging cy trong tp on ging ch c mt vi c tnh tt. Do vy, mun c 1 ging cy trng l tng cha ng tt c cc gene tt ca nhiu ging phi tn thi gian mi lm c. Vic tng hp gene mang c tnh tt ca cy trng thng c lm bng phng php lai v phi
lai trn nhiu cp phi hp vi nhau tng i mt mi mong t c kt qu tt.Ngi ta cn p dng u th lai i vi cy th phn cho (th phn khng phi ht phn ca mnh m ca cc cy khc qua vic to dng thun chng 6-7 th h.V d, ng t lai n sang lai kp, lai 3: 1 cp lai n lai vi 1 dng t phi. Sau ny cc nh khoa hc pht hin rng b v m c c tnh tt th con lai cng li ci tt ca b v m v tt hn b m. l tc dng cng ca gene v nu con lai tha hng cc c tnh tt ca nhiu b v nhiu m th con lai cng tt hn - gi l lai tng hp. 1.3. t bin De Vrie (1901) nghin cu tnh t bin thc vt v nhn ra tnh v hng ca n.ng tch ri t bin vi ngoi cnh v i n ph nhn tc dng tch ly bin d ca chn lc t nhin. W. Johnson (1903) chng minh ch c bin i trong gene mi di truyn c (t bin). Bin d t bin l do s bin i vt cht di truyn (nhim sc thNST, gene) gy nn. C 3 loi t bin: t bin gene, t bin nhim sc th v t bin gene t bo cht. * t bin gene hay t bin im l nhng bin i gene xut hin mt cch ngu nhin hoc nhn to. l s thay i cu trc ca gene xy ra theo cc kiu: mt i mt cp nucleotid, thm vo mt cp nucleotid v bin i trnh t cc nucleotid. Bin i gene thng c hi, ch rt t trong trng hp c li, s c dng lm nguyn liu cho qu trnh tin ha. * t bin nhim sc th l loi t bin mi c th. C 2 loi t bin nhim sc th: t bin s lng v t bin cu trc nhim sc th. - t bin s lng NST l nhng loi t bin v s lng NST ca mi c th. Nhng t bin s lng c th xy ra ton b NST (t bin a bi) hay mt cp NST no . t bin a bi c th to ra c th c b NST tng ln 3n, 4n, 5n,..NST. t bin mt cp NST v d cp NST th 21 ngi c 3 chic gy bnh Down, t bin tng s NST gii tnh gy cc bnh v sinh. - t bin cu trc NST l nhng t bin trong cu trc b NST. Nhng t bin v cu trc NST c th xy ra cc dng sau: . Mt on: mt i mt on no ca NST. . o on: hai phn trn cng mt NST o v tr cho nhau. . Thm on: gn thm mt on mi vo NST. . Chuyn on: s trao i hai on trn hai NST khng cng ngun cho nhau. * t bin gene t bo cht. C nhiu trng hp kiu gene bnh thng m c bin i kiu hnh. l do bin i gene t bo cht gy
nn. Cc t bin gene t bo cht c nhng c im ging t bin gene trong NST, chng cng bn vng v di truyn c cho i sau. t bin gene t bo cht c th ln hoc tri. t bin gene t bo cht cng l ngun nguyn liu cho chn ging. Nguyn nhn ca s t bin l do tc nhn bn trong t bo v bn ngoi mi trng gy nn. Cc yu t bn trong t bo nh qu trnh trao i cht, hot ng sinh l mt bnh thng c th gy t bin. Cc yu t bn ngoi nh tc nhn vt l: nhit cao, siu m, tia phng x, tia t ngoi, hay tc nhn ha hc, c bit l colchicine, l cc tc nhn gy t bin quan trng. Con ngi c th s dng n to ra cc t bin mong mun dng cho chn ging. Nhng vn nu trn tuy gi l CNSH truyn thng nhng thc cht cho n ngy nay ngi ta vn s dng n trong vic to ging mi.
2. CNSH trong vic ci to ging v pht trin cy trng cho nng sut cao
T nhng nm 1960 tip sau vic du nhp cc ging mi vo chu v chu M Latinh, danh t cuc cch mng xanh bao trm tt c nhng c gng tng nng sut nng nghip cc nc ang pht trin (PT) thng qua ging mi cao sn c bit l la v la m. S dng cc ging mi ny i hi nhiu loi thuc tr su, cc bin php ti, phn bn v chm sc. Lai cc ging mi ny vi cc ging chng chu ca a phng cho php thu c cc ging c nng sut cn cao hn v thch nghi tt hn. ng thi, ngoi la v la m, cng tc nghin cu c m rng ra k, la min, ng, Triticale v mt s cy u (Sasson, 1986). Ch trong vng hn 10 nm, mt na din tch la m v 1/3 din tch ng la ca cc nc PT gieo bng ging mi cao sn. Nu c ti nc hp l, bn phn v x l thuc tr su, nng sut c th tng gp i n gp ba ln ging a phng. Cuc cch mng xanh ln th hai c ni n vo nhng nm 1970 l kt qu cc cng trnh nghin cu lai to ging nng sut cao, chng chu su bnh, hn hn, c th pht trin trong iu kin t phn bn v thuc tr su hn. Cc cng trnh nghin cu ny s khng cn ch da trn cc k thut lai, th phn cho m dng k thut nui cy m v t bo cng vi k thut ti t hp di truyn nhn nhanh cc ging ng ch v to ra ging mi (Murashige, 1974; Vasil, 1984;, 1985, 1986,; Arnon, 1985; Mantel et al., 1985; Zaithin et al., 1986; Davies, 1097; Picrik, 1987,), cc k thut ny da trn cc c ch t bo hc v sinh hc phn t quyt nh tnh a dng sinh hc (Collins, 1982, 1984; Kosuge et al., 1983; Tudge, 1983; Gelvin v Schilperoot, 1988).
Nh trnh c vic lai cho v vt qua c tr ngi ca tnh khng tng hp sinh dc (sexual incompatibility) nn tit kim c nhiu thi gian. K thut ti t hp DNA v ng dng chng dn dn i vi thc vt c th gip loi b hng ro sinh l v gii phu ngn cn s lai khc loi (Rachie v Lyman, 1981), cc k thut in vitro cng cho php tng s a dng di truyn gn y b gim st do s ph hy cc sinh cnh t nhin, lm gim s e da do su bnh mt s cy trng c nn di truyn qu n thun. Cc nc ang pht trin nht l cc nc trong khu vc nhit i cn ang gi c s a dng di truyn tng i rng trong cc h sinh thi t nhin v h sinh thi nng nghip ca mnh di hnh thc nhiu loi hoang di c quan h h hng vi cy trng, nhiu ging chng chu v ging a phng. Cc k thut nui cy c quan, m t bo thc vt v cc lnh vc nghin cu lin quan c th lin kt vic p dng cng nghip theo cc con ng sau: Nui cy nh sinh trng v ti sinh cy hon chnh Nui cy t bo trn protoplast, m n bi, chn lc cc bin chng v t bin, dung hp protoplast v ti sinh cy K thut ti t hp di truyn, cy chuyn gene v ti sinh cy. Nui cy t bo qui m ln, chn t bin, dung hp protoplast v k thut ti t hp DNA. Nui cy t bo, protoplast, chuyn dng sinh hc. Nhn ging in vitro cc cy v cc ging sch virus Ci thin ging v nhn ging cy trng Ci thin ging cy trng. Sn xut cc loi hot cht c ch. Tng hp cc cht mi.
Trn quan im kinh t, chi ph lao ng tham gia vo cy chuyn nhn ging l khon mc ln nht trong gi thnh ca cy ging in vitro. Ngi ta ang tin hnh nghin cu cc my t ng cy chuyn. Nm 1985, mt mu my cy chuyn c ch to ti Australia c th cy c mt cy trong mt giy. Nhng thit b nh vy c th lm gi thnh cy cy m gim i rt ng k (Marti, 1986-87). Vic p dng k thut in vitro c th c xp thnh 3 loi:
. Cc p dng c c trong thi gian ngn (3 nm): nhn ging v tnh in vitro, sn xut cy sch bnh, bo qun v trao i qu gene thc vt. . Cc p dng trung hn (3-8 nm): cc t bin soma v t bin giao t, cu phi, th tinh trong ng nghim, nui cy ti phn v sn xut cy n bi. . Cc p dng di hn (8-15 nm): lai t bo soma, lai xa, dng t bo t bin, chuyn gene, chuyn NST, sn xut cc cht th cp bng t bo nui cy in vitro. Vic p dng CNSH vo i mi thu hoch ma mng ang l mt trong trong nhng cuc cch mng khoa hc v k thut hin nay. Vic i mi ny c th da vo cc bin php k thut chnh sau: * Sn xut nhanh v qui m ln nhng cy trng c cng mt tnh cht di truyn, cho nng sut cao thng qua k thut nui cy m v t bo. * To nhng ging mi c nng sut cao thng qua phng php chn dng soma trong nui cy m t bo. * To ra nhng cy lai mi c c tnh u vit bng k thut dung hp protoplast (protoplast fusion). * To ra nhng c tnh mi mong mun qua vic a cc nguyn liu di truyn vo t bo cy trng bng k thut ti t hp DNA 2.1. Sn xut nhanh v qui m ln nhng cy trng cng c tnh cht di truyn cho nng sut cao thng qua k thut nui cy m v t bo Trong k thut trng trt c nhiu loi cy cn phi nhn ging v tnh qui m ln. Mt s cy trng c th ti sn xut d dng bng ht nhng kh nng ny mm thp, c bit l cc cy lm nghip. Mt s khc tuy ht d ny mm nhng qu trnh sn xut ht li qu t. Cng c mt s cy lai duy nht cn c nhn ln v tnh gi li nhng c tnh u vit. Trong nhng nm 1930, vic ti sinh li chi v ton b cy trng c tin hnh mt cch thun li nh xy dng c k thut nui cy m v t bo thnh cng. Ngy nay, hu ht phng th nghim nghin cu sinh l ha sinh di truyn thc vt u c trang b k thut nui cy m v t bo. K thut nui cy m v t bo thc vt bao gm: * Chun b mi trng nui cy c thnh phn cn thit v nhiu loi, cng nh phi chn ging ng cho mi trng nui cy tng loi m, t bo v thay i theo tng thi k pht trin v phn ha ca m (mi trng nui cy protoplast khc vi mi trng nui cy callus, mi trng to r to mm khc vi mi trng duy tr m trng thi callus, v.v.)
* iu kin v trng phi nghim ngt, k t khi chun b mi trng n khi x l m. V vy, phi c bung cy v trng v t cy Laminaire, cng nh cc thao tc dng c u phi tun theo nguyn tc v trng trit . * Chn la m phi c iu kin pht trin mnh v phi c kh nng to thnh callus trong mi trng cha cht dinh dng thch hp. Thng ngi ta chn m phn sinh ngn hay chi nch. * iu kin x l m phi thch hp. Tuy cc m trn cng mt cy cng mt lng thng tin di truyn nh nhau nhng cho cc callus pht trin hon ton khc nhau trong kh nng ti sinh chi, pht trin r hay thnh cy hon chnh. l do x l cht iu ha sinh trng (HST) khc nhau, x l nhit v nh sng khc nhau. Phng php nui cy m v t bo thc vt l phng php nhn ging l tng khng ch do i hi t din tch, nhanh, m cn gi nguyn c tnh u vit ca ging cy ban u. Nhn ging v nhn dng v tnh c ngha c bit i vi cy nhit i v chng c d hp cao, thng nhim nhiu loi virus. Cc cy trng sau u c th a vo nhn ging v tnh in vitro vi mc tiu thng mi ha trn qui m ln: Atiso, mng ty, c ci ng, ti, gng, khoai ty, Raspberry, du ty, ma ng, khoai lang, khoai nc, da di, hnh nhn, to ty, chui, cam, chanh, da, anh o, kiwi, c du, u , l, da, chui bt, nho, ht d, tre, lim, bch n, v, cm chng, cc, Iris, Gerbera, hu, lan, Pelagonium, quyn, hoa hng, Mt s cy ang c ti sinh trong phng th nghim: cy b, ca cao, c ph, Jojoba, cao su, ch l, thuc l, c rt, Endive, ci du, ng, u, c t, u nnh, (theo ti liu Zimmerman, 1986; Ketchum, 1987; Picrik, 1987). Trung M v Nam M, k thut nui cy m c p dng nhm to ging cy sch bnh v nhn ging v tnh cy c du (Brazil, Colombia, Costa Rica, Cng ha Dominique), cam, chanh, khoai ty, du ty (Brazil), c ph (Costa Rica v Mexico). Nhiu cng ti t nhn cng dng k thut ny tng sn lng c du (Costa Rica, Cng ha Dominique), chui (Honduras), lan (Brazil) cm chng, cc, da cnh (Colombia, Costa Rica). Nm 1987, khoa Sinh hc i hc Maranho thnh lp mt phng th nghim cy m thc hin chng trnh chn ging cc cy n qu nhit i: da ht, v cc cy g cung cp lng thc khc. Cc cng ti t nhn Brazil Biomatris S.A. (Rio de Janeo) l chi nhnh ca cng ti ging khng l AGROCERES ang tham gia vo cc nghin cu trin khai vic nhn ging in vitro khoai ty, cy n qu n i
v nhit i, cy cnh. Cng ti tri rng trn ton lnh th Brazil v sn lng hng nm ca n ti 2,4 triu cy ging (Biotechnologia Fundaco). Vit Nam, Trung tm Thc nghim Sinh hc ti thnh ph H Ch Minh (1979-1980) cng nhn ging v tnh in vitro ging khoai ty phc v cho cc hp tc x sn xut thnh ph Lt. Vin Khoa hc Vit Nam H Ni (nay l Vin Khoa hc v Cng ngh Vit Nam) cng th nghim nhn ging v tnh in vitro cc cy khoai ty, c, la, thuc l t nm 1974-1975. Cho n nay, y cng nhn nhiu ging cy trng nh ma, ng, da si, la, thuc l, c kh nng chng chu phc v cho vic trng trt a bn min Bc. i hc Nng nghip I, vin Di truyn Nng nghip TW, cng bng nhn ging v tnh v k thut dung hp protoplast to ra nhiu ging cy trng phc v cho sn xut nng nghip. Vic nhn ging v khai thc cy chu hn (serophyte) mang li nhiu mi li cho cc nc PT vng kh hn hoc bn kh hn. Trong s 350.000 loi thc vt c cc nh thc vt hc m t, con ngi mi ch th trng khong 3.000 loi lm lng thc, ly si, lm thuc hoc thu nguyn liu. Ch c khong 100 loi c trng din rng v 90% lng thc ca loi ngi do khong 10 loi cung cp, trong khng c loi no thuc cy chu hn. V vy, cn thit phi tm ra cc loi cy chu hn c kh nng cho sn phm di do cc vng kh hn chim hn 1/3 din tch ca qu t. Cc ngun nc ti ngy nay ang tr thnh mt nhn t hn ch trong s pht trin ca nng nghip. V vy, tm cy chu hn c ngha quan trng trong sn xut. Nm 1960, Vin Nghin cu ng dng, i hc Ben. Gurion Negev, Israel c thnh lp vi mc ch du nhp v pht trin cc cy thch nghi vi iu kin kh hn v bn kh hn. Lc u vin thc hin ci gi l nng nghip sa mc ngha l du nhp v pht trin nhng cy t vng kh cn, cc loi s dng rt t nc ma (lng ma di 200 mm), ch cn b sung nc ti thiu. Sau , cc nh khoa hc Israel chuyn sang lm nng nghip trn sa mc, ngha l lm cho nhng ngi nh c trn vng kh cn c thu nhp cao cho h c mc sng kh. Ngi ta a vo s dng vic ti nc l hay mn (nc ny c vng sa mc Negev). Vin Rodolph v Rhoda Boyko (Vin Nghin cu Nng nghip v Sinh hc ng dng) ca Israel tin hnh nhiu chng trnh nghin cu nhm p dng cc tin b nng hc v CNSH vo vng sa mc Negev v cc vng kh hn ni chung (chng trnh c s tham gia ca Israel, M, Ai Cp, H Lan, Cng ha Lin bang c) theo ti liu ca Raz, 1987). Ngi ta trng nhng cy chu hn nhiu nm trong c cy cao v
cy bi Atriplex mummularia (Saltbusch) Atriplex canescens v Cassia sturtii) cho cc kt qu c bit tt. Qua nghin cu so snh 120 loi cy chu hn th Atriplex nummularia, Atriplex barclayama v Atriplex lentiformis l cy chu mn cho nng sut cao v dng lm thc n gia sc. Cy Distchlis spicata (c chu mn) cng c th sng trong iu kin cc kh hoc mn dng ph xanh v ci thin nhim vng Texcoco (Mexico). Cy Jojoba (Simmondsia chinensis) l loi cy bi c l thng xuyn thuc h Buxaceae (cao n 5 m) tm thy ty bc Mexico trong sa mc Sonora v c vng kh cn bang California v Arizona ca M (c th mc sa mc c lng nc ma 75 mm vn cho qu tuy cy c thp). Cy Jojoba c b l dy, th, chu nhit 50oC nh b r n su 30 m. T xa xa, du Jojoba dng bi tc v x l da sc vt (th dn Apaches s dng). Ht Jojoba (bng ht Lc) cha mt loi sp lng chim 30-60% mu hi vng, c mi, thnh phn khng cha glyceride m cha mt hn hp cc ru v ester ca cc acid bo mch di t 2022 nguyn t C. Du Jojoba thay th du c voi dng bi trn trc chuyn thy lc v hp s xe ua p sut v nhit cao, dng trong cng nghip da, cng nghip m phm, cng nghip dc, cht chng bt ln men vi sinh vt, sp bng ph cc loi giy carbon c bit. Kh du cha du d v khong 30% protein, x, tannin v cc cht khc. Cy Guayule (Parthenium argentatum) l cy ly nha m t nhin lm cao su. Cy Crambe (Crambe abyssinia) thuc h Thp t Cruciferae cha mt lng ln acid erucic c th thay th ci du. Cy b tru Cucurbita foetidissima (Buffalo gourd) c ht giu du v protein, r cha nhiu tinh bt. Sau 4-5 nm sinh trng, thn, l, r nng 40 kg trong c 20% l tinh bt, chi Grindelia gm nhiu loi dng lm nha do. Cy Ocnothera spp. l cy lm thuc, ht c nhiu acid -linoleic c dng nh cht b sung dinh dng v lm m phm. Nhng cy nu trn, ngi ta dng CNSH nui cy m v t bo nhn ging v trng qui m rng, va chu hn, chu mn, chu nng, chu ngho dinh dng m t nng sut cao v dng trong nhiu ngnh cng nghip khc nhau, phc v cho i sng. i vi cy rng, xut khu g gi vai tr quan trng i vi cc nc PT. Theo s liu thng k ca b Nng nghip Php: nm 1984-85, mu dch g nhit i l 35.236 triu m3 trong chu Phi: 35%, chu : 60%, Trung v Nam M: 5%.
Nhn ging v tnh in vitro cc cy rng ly g hay lm bt giy c ngha kinh t rt ln. Chi bch n (Eucalyptus) c nhiu loi c hu Australia, Timor, Tn Guin, Philippinnes. Bch n du nhp v trng Nam M, chu Phi, Spain, Portugan, chu , Trung Cn ng v Bc M. Cc phng php nhn ging v tnh truyn thng nh gim cnh, chit cnh, ghp i vi bch n u khng cho hiu qu. Ngi ta to callus t nhng phn khc nhau ca cc loi bch n v cy con ti sinh t callus t cc b phn khc nhau ca bch n chanh Ecalyptus citriodova v bch n trng E. alba. T nm 1970, nui cy thnh cng mnh l, on thn, r bch n. Cc nh nghin cu M thu nhn cy con nui cy on thn cc loi: E. grandis, E. gunni, E. dalrrympleana, E. pauciflora, E. ficifolia. T nm 1973, AFOCEL (Association Franaise Fort-cellulose) khi s nhn v tnh in vitro cy bch n nhm mc tiu sn xut ln cc dng v tnh chu lnh v nng sut g cao. T nm 1975, bt u trng ngoi t c mi thng trng 20.000 cy bao gm 18 dng v tnh. Hartney (1982) nhn v tnh thnh cng cc ging E. camadulensis, E. curtisi, E. ficifolia, E. grandis, E. obtusifolia v E. rudis, bng cch nui cy chi nch v t cy con.Mchra-Palta (1982) thnh cng trong to chi ph t l mm v trn on thn bch n E. nova angelica v E.viminalis trong iu kin in vitro. Diallo v Duhoux (1984) lm vic ti phng th nghim Sinh l thc vt i hc Dacar, Senegal nui cy l mm v to thnh cng chi cy E. camaldulensis, trn mi trng c cha NAA (naphtalen acetic acid l mt auxin) v 6.BA (6-benzylaminopurine l mt loi cytokinine) k thut ny cho php to ra 200 cy t 1 cy con trong 2 thng v 1013 cy trong 1 nm, trong khi k thut ct on ca Gupta ch t 106 cy/nm. Davies (1984) lm vic ti phng th nghim ca Dhoux pht trin k thut nhn v tnh cy Faidherbia (Acacia albida). Cy h u ny mc hu ht cc vng kh hn chu Phi, c bit l Ty Phi. Chng ng vai tr quan trng i vi kinh t ng c vng Sudan Sahel. Chng c nh N2 v rng l vo ma ma. L v qu dng lm thc n cho i gia sc vo ma kh. Acacia albida to mi trng thun li cho cc cy k, la min, lc mc di tn l cy ca chng vo ma ma. Hai cy rng khc c kh nng c nh N2: cy h u Acacia senegalensis v cy khng h u Casuarina equisetifolia cng c quan tm nghin cu nui cy m.
Nghin cu to phi soma Mt hng khc c t chc trng trt l vic to phi soma. Theo nh m t ca Steward v cng s, s chuyn sang mi trng c nng auxin thp gy ra s sinh trng t bo trong phi. Chng cng tri qua tt c cc giai on pht trin bo thai ca mt hp t nhng to thnh t t bo soma ch khng phi l sn phm ha hp ca 2 giao t c v ci. S nui cy phi ca t bo soma c mt s tin b: * Phi pht trin c hai hng em n c r, chi v pht trin thnh cy ton vn ngay t u. * Nui cy phi c th to ra mt hng ln cc cy hn c con ng nui cy m. * Khi ln ln trong mi trng nc th phi tch ra thnh nhng phi khc v bi t do, do , khng cn nhiu thit b. Hng ngn phi pht trin trong bnh nui cy c tht v cng c tc sinh trng nhanh khng km tc sinh trng khi nui cy vi sinh vt. Ngoi c rt, cn ty, nhng cy khc nh u, chanh, c ph, ch l, k cng c nui cy phi t bo soma. S chn phi t bo soma c th c ci bin bng nhng cht HST c bit dng acid abscicic v thay i mi trng. Cng ngh hin nay hon ton cho php nhn ln nhng phi soma cho mt s ln cc loi cy thu hoch quan trng. Mt vi cy trng c ti sinh thnh cng bng nui cy phi soma: c rt, cn ty, u, c ph, chanh, c du, v.v. Th d: c du to t phi soma c th thc hin trc tip t callus s cp trn mi trng cha auxin, cytokinine v than hot tnh. Cc th phi hnh thnh 6 thng sau khi a cc phn on ca l v nui cy. C th thu c ti 500.000 th phi t 1 mu l trong vng 1 nm. Cc th phi thnh thc, c tch ra v to cy, cc phi non dng tip tc nhn (Noirel, 1984-85). Chng ra c th phn bit 2 giai on trong nhn ging in vitro c du: giai on th nht: t lc to callus rt ngn cn 6 thng hay t nht ty tc to phi soma, giai on hai: thng ko di 4 thng trong phi t nhn ln v hnh thnh cy. Vic chuyn vn th phi t Php n Malaysia hoc Indonesia khng c g kh khn. Gi ng lnh th phi c thc hin ti phng th nghim Nghin cu Sinh l cc c quan thc vt Sau thu hoch CPOVAR ca vin Nghin cu Khoa hc Quc gia Php CNRS. Phi c bo qun -196oC trong thi gian rt di. Ngi ta nhn thy rng cc cy c du xut x t cc phi ng lnh sau 15 thng trng trn i khng c bin d hnh thi no. V vy, Vin Nghin cu du v cy c du Php (IRHO) xut k thut ny cho cc khch
hng hi ngoi. K thut ny rt thch hp i vi cc loi khng thch hp vi nhit bo qun ca cc cy n i (0-5oC). Hn na n c th gim thiu cc him ha tim tng nu s ln cy chuyn nhn ging tng ln qu ln (Biofuture, 1987, s 56). Du c l loi du n c s dng nhiu nht trn th gii, ch ng hng th 2 sau du u tng v chim 17% tng sn lng du bo v m (3-11 tn/ha/nm), nn cn c ging c du trng, nht l cc nc cn xch o. V cy da, t nm 1981 cc nh khoa hc Php thuc Vin Nghin cu Khoa hc v S pht trin (ORSTOM) v IRHO cng bt u cc th nghim nui cy m da nhm to phi soma. Cc cy nui cy m c to t m ca phn l non ca cc cy da 5 tui trng trong nh knh thuc ging PB-121 con lai gia ging ln vng Malaysia (Malaysia Yellow Drafs) v ging cao Ty Phi c chn lai ti Cte dIvoire. Cc th phi thu c sau 6 thng nui cy. Cc nghin cu to cy hon chnh t phi c tin hnh t nm 1984 (Pannetier v Noirel, 1984). Theo cc nh khoa hc Php, qu trnh pht sinh phi soma c th xut hin trong mt giai on ngn t m l non ca cy da t 2 n 5 tui v c th thu c vi cy trng thnh . Bng phng php ny, da cho nng sut cao hn (s qu v sn lng cm da) c th gp 5 ln. i vi cy c ph, cc nh nghin cu Php GERDAT (Groupement dEtude et de Recherche dAgronomie Tropical), Montpellier nui cy m c ph t nm 1978-1979 v nm 1981 thnh cng trong thu nhn phi soma trc tip t m cy ban u m khng phi thng qua giai on to callus trung gian (Dublin, 1982). Cc phi v tnh c hnh thnh t cc b mch ca phin l em nui cy trn b mt mi trng. T mt mnh l duy nht c th to ra hn 1.000 cy con. Qua mt vi v d c th nu trn cho thy rng, vic nghin cu to phi soma l mt trong cc bin php k thut c trin vng ca CNSH, nhm nhn nhanh cc ging cy trng nng lm nghip qui m rng. Nghin cu nhn ging cy sch virus. tin hnh to cy sch bnh virus bng k thut nui cy m v t bo, ngi ta thng dng m phn sinh nh chi. Nhng m ny cha nhng t bo sinh trng v c bao mt lp v cutin. S hnh thnh mi cc c quan ca thc vt bt u trong cc m phn sinh nh chi ny. Cc m phn ha ngay t nhng giai on u ca phi v gi li trong sut qu trnh sng ca cy. M phn sinh l vng khe mnh nht ca cy, v ngi ta thy rng, qu trnh sinh tng hp DNA ca virus
thc vt khng xy ra c vng ny do mt c ch g hin nay cha r, thm ch c cy b bnh virus nhng phn ny vn khng b nhim virus. V m phn sinh c cc t bo phi nn khi nui cy to nn callus. Dng cc cht HST khc nhau nh gibbrelline, auxin, cytokinine, v.v. kch thch khi t bo khng phn ha ny m chi. T hnh thnh nn nhng cy con khe mnh khng b virus. Bng cch ngi ta sn xut ra du ty, khoai ty, u , khoai m, sn v nhiu cy cnh sch bnh virus. Nhn ging bng sn xut ht nhn to. Ngy nay ngi ta dng k thut nui cy m v t bo thc vt sn xut ht nhn to. T bo thc vt c c trng l khng ch tr thnh t bo sinh dng m cn tr thnh t bo phi mm, ch cn iu khin chng bng cc cht HST thch hp. Do vy, ngi ta cho vo bnh dung dch dinh dng c cht HST nht nh th t bo c th tr thnh t bo phi. Cc t bo phi trong bnh s sinh sn rt nhiu v t hp li. Cc phi ny c bao bc bi mt cht keo - gm hn hp cc cht dinh dng v gi l cc ht nhn to. Khi gieo cc ht ny xung t s mc thnh cy bnh thng. Vic sn xut cc ht nhn to nh th rt cn thit trong nng lm nghip. Bi v, trong ci to ging nhiu khi to ra c ging c nng sut cao v chng chu gii nhng li khng c ht hoc rt t ht c th gieo trng li. Bng phng php ni trn, ngi ta c th sn xut ht nhn to bng cc t bo bnh thng ca cy ny vi mt lng ln trong cc ni ln men. Mt lot cc vn l th v ht nhn to y l: * Khi cn trong cc bnh nui cy ln men rt tin li cho ngi ta x l nhit cc m lm sch ht virus, to ra cc ht sch bnh. * Ngi ta cng c th a vo v ht nhn to cc loi vi khun c nh N2 th khi cy trng thnh, vi khun ny s ly N 2 t khng kh cung cp phn m cho cy . * Cng bng cch tng t nh trn, ngi ta a mt lng thuc tr su hoc tr c di vo v ht nhn to bo v cy khi b su v c di ph hoi. * Ht nhn to cng l i tng d dng np cc gene l vo to ra cc ging mi c c tnh mong mun. Rt mng l hin nay trn th trng th gii c bn nhiu ht nhn to vi cc tnh u vit ni trn, trong c ht la m, ht la, l nhng ht lng thc sng cn i vi cuc sng con ngi. 2.2. To ging mi c nng sut cao thng qua phng php to dng soma trong nui cy m t bo
Soma l tn gi cc t bo c th (sinh dng), n khc vi t bo sinh dc. Chng ta bit rng, t cc t bo soma c th to nn bt k b phn no ca cy. l c im ton nng ca t bo thc vt. T callus c th khi phc li mt b phn no : r, thn l hoc to thnh mt cy hon chnh u da trn c im ton nng ny. C iu callus c cy i cy li nhiu ln thng xy ra nhng bin i di truyn. Nhng cy ln ln t nhng t bo bin i y cho cc ht. Nhng ht ny li mc thnh cy cho nhng c tnh qu m ta mong mun. Nh vy, chnh lc ny nhng cy s l ngun ban u nhn ln v nhng th h sau c to ra m ta gi l cc dng soma. Phng php to ging kiu ny gi l phng php to dng soma. C s khoa hc ca vic chn ging l hin tng bin d soma - tc l bin i di truyn khng phi t bo sinh dc m t bo c th (sinh dng) ca cy. C ba cch: * Bin i NST kiu a bi th (polyploid) nh c chua, u, cy cnh. * Bin i NST kiu thm, bt mt vi NST trong b NST t bo. * Bin i kiu t bin mt s gene nht nh NST hoc gene ca ti th, lp th (lc lp). C nhiu kiu bin i: . S bin d t nhin: thng rt thp, ch mt trong hng triu t bo. . Bng bin php chn lc c in (sau lai ghp v t bin) th kh c c s phi hp gia ci c v ci mi. . Bin i dng t bo soma, c bit l t nhng t bo callus trong nui cy th tn s xy ra rt nhiu. Khoai ty ti sinh t callus c th nghim, trong c 13 bin d gene n gin c pht hin trong 230 cy khoai ty ti sinh. T l bin i dng soma y l 1/18, c ngha l t 18 cy ti sinh th c mt cy bin d. T l bin d ny tht qu ln. Mt s bin d gene n gin xy ra c chua cng c xc nh v mt trong nhng bin d nm cnh tay di ca NST 10 trong bn gene. Va qua t c mt s kt qu v bin d soma trong s cy trng kch thch s p dng phng php chn lc dng soma vo vic i mi cy trng. Bng phng php chn dng soma, ngi ta to nn c ging la va chn sm va c ht dng di nhm va tng nng sut thu hoch li va to c cht xanh lm thc n cho gia sc.
V cy ma, phng php chn dng soma ny to ra c ging ma chng c mt s bnh virus nh bnh fip, nm lng, bnh than, v bnh t mt. Ta bit rng cy Ma l cy bt bi (octoploid) hoc thp bi (decaploid) v rt d hp t. iu ny gy tr ngi cho vic m hnh ha l thuyt v cc phn tch di truyn. Heinz, 1977 Hawaii v sau cc nh khoa hc i Loan, Php, Cuba, Argenetina, thnh cng trong ti sinh ma t callus, t nui cy ti phn to c cy n bi c nng sut cao hn trong iu kin t nhin so vi cy b m, hoc phng php chn lc dng soma cho php phn lp c cc dng khng bnh. Bng phng php chn dng soma, cc nh nghin cu cng thc nghim chng bnh tn li cy c chua, chng virus khm thuc l trn cy thuc l. T nhng bin i dng soma, cng pht hin nhng qu to nh khc nhau, t p dng vo vic chn lc cy n qu nh bi, cam, thu c nhng c tnh qu. i vi cy c du, khi ti sinh t callus, cy khng ging nhau m xut hin mt s t bin soma (Jones, 1983). Nhng bin d soma ny theo Cocking i hc Tng hp Notlingham, c nguyn nhn di truyn v do c iu kin nui cy to nn (Cocking, 1981). Ni tm li, trong nui cy m v t bo thc vt, ngi ta tha nhn c s bin i di truyn. S bin i nh vy c th c ch i vi thu hoch ca con ngi. Trong qu trnh nghin cu c s di truyn ca s bin i t bo thc vt nui cy, ngi ta nhn thy rng, cng ngh nui cy t bo in vitro cho chng ta mt li kh quan trng lm thay i ging cy trng theo hng c li. l nhng bin i dng soma gip ta to ra nhiu ging cy mi vi c tnh u vit. 2.3. To ra nhng cy lai mi c tnh u vit bng k thut protoplast Protoplast l t bo trn. Thnh t bo cellulose b tiu hy bi enzyme cellulase hay drilselase, n ch cn mng sinh cht bao quanh. Protoplast c kh nng dung hp (fusion) vi nhau to thnh cc th lai v tnh di tc dng ca mt tc nhn ha hc hay vt l lm cc mng kt dnh li vi nhau v gy ra s chuyn vn cc i phn t DNA, cc protein, cc bo t t t bo n sang t bo kia. Dung hp l qu trnh hp nht 2 protoplast li lm mt. Hin tng ny ph bin. S tht con ngi sinh ra cng l kt qu mt s dung hp gia tinh trng v non trng, to thnh hp t c du hiu di truyn ca c 2 t bo hp li. Ch c iu qu trnh gi l lai hu tnh. Cc t bo thc vt c kh nng dung hp di dng protoplast. Hai protoplast
dung hp li c th sinh ra mt cy con hon chnh di du hiu di truyn ca hai t bo hp li. Nhng qu trnh ny l qu trnh lai v tnh (somatic hybridization). Ngy nay nh lai v tnh m ngi ta ci to c nhanh chng b my di truyn t bo nhm to nhng ging mi c nng sut cao. S dng k thut protoplast, ngi ta lai to gia cy trng vi cy hoang di khng mt s trng thi trong mi trng bt thun nh to ra nhng cy chu hn cao, chu rt gii, chng chu su bnh, phn, mn, v.v. K thut nui cy v dung hp protoplast n nay khng nhng thc hin tt i vi cy 2 l mm m ngay cy c mt l mm nh la, thm ch cc loi rong, to. K thut protoplast trong to ging mi ca thc vt gm 2 giai on chnh sau: a. Giai on tch v nui cy protoplast. giai on ny ngi ta c th tch t bo x l to thnh protoplast t tt c cc thnh phn ca cy nh r, thn l, nt sn r, l mm, ht phn, callus v.v. nhng hay dng c l t m l. Giai on ny gm cc bc sau y: + X l mu cho sch v v trng bng cn 70o, sau bng calcium hypochloride, cui cng bng nc ct v trng. + Ct mu. Mu c ct nh thnh mnh 0,52 cm hoc thnh si. + X l mu bng enzyme. S dng enzyme hn hp nh pectinase, cellulase. Onozuka R10 trong mannitol pH=5,8 mt thi gian trong iu kin nh sng v nhit ti u. + Tch, lm sch protoplast bng phng php lc v li tm. Cc bc chung l nh vy, song chi tit mi loi cy i hi mt loi enzyme tch protoplast, nng enzyme cng nh nng cht gy co nguyn sinh cht khc nhau c lng t bo protoplast cao nht. b. Giai on dung hp protoplast. Trong giai on ny ngi ta s dng mt trong hai phng php sau y dung hp: * Phng php s dng tc nhn ha hc: s dng cht polyethylene glycol (PEG). Cht ny gy nn kt dnh 2 protoplast v to iu kin cc phn t DNA cng nh cc thnh phn khc d i qua mng. * Phng php s dng tc nhn vt l: cc xung in (electroporation) to nhng l nh ca mng, to iu kin cho cc i phn t i qua. Thc cht nhng vn chnh ca k thut dung hp
protoplast l vic phn bit v tch th lai thng qua mt thng, dng gene nh du, dng mi trng chn lc, dng phn ng mu enzyme, dng protoplast t cc loi m khc nhau, hoc c nhum bng cc cht pht hunh quang. Qu trnh dung hp c th thc hin cc bc sau: + Hai t bo protoplast sch ca 2 loi cy nh lai cho tip xc nhau theo t l nht nh ri nh cc git dch hn hp ln a nha trong petri. + Nh nh nhng PEG ln nh mi git dch cha protoplast hn hp , mt thi gian nhit phng ri nhc li vi ln nh th. + Ra sch PEG v cho vo mi trng nui cy c cha mui ammonium phc hi t bo sau qu trnh tch v dung hp. + vi ngy sau, t bo phn chia, cy chuyn sang mi trng chn lc v nh vy cy chuyn mt vi ln na trong mi trng cha nhng cht HST thch hp to callus, to chi, to cy v r,v.v. + C th th phn ng enzyme trn mu l cy lai ct nh c th nhn ra c c s lai to hay khng. Ngy nay ngi ta ang s dng k thut dung hp protoplast to ra nhng cy lai mi. Chng hn, lai khoai ty trng v khoai ty hoang di. Gene cy hoang di gip cy trng chng bnh virus.. Ngi ta lai cy c chua vi cy khoai ty (h hng chng xa nhau) cho cy lai mang qu c chua v c khoai ty. Bng cch ny, ngi ta to ra cy lai gia c rt v rau mi, cam v chanh, i vi c du, t protoplast v s dng k thut ti t hp gene c th nng cao hiu qu, chn lc cc dng c du c nng sut cao.
Hnh II.1. Cy Khoai-C (Pomato)
i vi ma, dung hp protoplast c ngha ln i vi vic ph bin cc ging ma khng ra hoa (trn c) hoc bt th. Cc kt qu nghin cu ca CENA (Centre de Nuclear Energy in Agriculture) v CEBTEC (Diviso de Biotechnology de Plantas et Centro de Biotechnologia Agricola) Brazil cho thy, dung hp
protoplast cy ma c th gip chuyn gene khng thuc tr su c t dng ma ny qua dng ma khc.
Dung hp protoplast c th chn gip cc dng c ph khng bnh v cc c t do nm tit ra a vo mi trng nui cy nhm chn ra cc dng t bo khng c t cho c ph. Nm 1987, Costa Rica cng ti sinh cy c ph vi t protoplast c tch t phi v tnh hnh thnh t huyn ph t bo callus l. Sau khi nui cy protoplast vi ln trn mi trng cha 0,5 mg/l kinetine, 0,5 mg/l 2,4D. 0,5 mg/l NAA, cc tc gi thu c callus nh. Khi cy chuyn sang mi trng khng c cht sinh trng cc callus pht trin thnh phi v pht trin thnh cy hon chnh. Ngi ta cng dung hp protoplast cao su t cc t bo phi v tnh. n nay c hn 70 loi cy cng c sinh ra t protoplast. Khi dung hp protoplast, ngi ta cn to ra c ging lai gia t bo thc vt v vi khun. S dung hp protoplast gia t bo to lam (va c c tnh quang hp va c c tnh c nh N2 trong khng kh) vi callus thc vt lm kch thch s pht trin ca chng. Cng vi k thut trn, ngi ta cn c th dung hp c cc t bo soma vi t bo sinh dc (ht phn) to ra cy lai gia cc h vi nhau. Tm li, k thut dung hp protoplast hin ti ang m ra mt kh nng rng ln trong vic to ta nhng ging c th tp hp c nhiu c tnh tt t cc cy khc nhau. Hn na, nh vic to thnh protoplast (bc trn v t bo), n tr thnh cng c nghin cu cc qu trnh sinh ha, c bit l acid nucleic v protein trong t bo, dng m hnh nghin cu di truyn phn t, nghin cu s xm nhp ca vi khun vo t bo, nghin cu qu trnh cng sinh vi khun trong cy b u, nghin cu cc enzyme, nghin cu c ch ca qu trnh quang hp v.v. lm c s khoa hc cho vic nghin cu ng dng phc v cho i sng ca con ngi. 2.4. To ra nhng c tnh mi mong mun qua vic a cc nguyn liu di truyn vo t bo cy trng bng k thut ti t hp DNA (DNA recombination) Vic ci tin a cc gene vo t bo thc vt qua cch a nguyn liu di truyn ngoi lai vo bng con ng ti t hp DNA hin nay ang m ra nhiu trin vng tt p: Cch th nht: chuyn gene trc tip . Phng php ha hc polyethylene glycol. . Phng php vt l xung in dung hp protoplast. . Phng php ngm ht phn vo dung dich DNA.
. Phng php vi tim gene. . Phng php bn gene. Cch th hai: chuyn gene gin tip qua s dng cc vector, c bit l vector plasmid pBI12A cha sn mt gene khi u promoter mnh nht v gene CaMV35S (Cauliflower mosaic virus) gene kt thc v gene nh du GUS-A. Plasmid ny c gn thm cc gene l c c tnh mong mun a vo vi khun hay thc vt dng Agrobacterium tumefaciene a vo cy.
Ngi ta c th a gene ngoi lai vo qua con ng vector plasmid: DNA c nh du phng x ri a vo ht phn, chi non, cc t bo nui cy protoplast hay nhn t bo c tch ra. DNA ngoi lai d nhn cm vi s tiu ha ca enzyme nuclease vt ch. Phi a DNA vo lyposome, sau dung hp vo protoplast lm gim tc dng tn cng ca nuclease t bo ch. Dn d ngi ta nghin cu a DNA ngoi lai vo qua con ng vector plasmid. DNA plasmid c tm thy ti th ca ng vt bc cao. Nhng plasmid c th l mt h thng duy nht ca s chuyn gene. Chng sao chp mt cch t qun trong ti th. Plasmid cng c tm thy trong nhiu vi khun. Nhng plasmid thng c lin kt vi vi khun bnh l v c dng kim tra s sinh bnh ca mt tc nhn gy bnh cy trng.
Vai tr plasmid Ti ca Agrobacterium tumefaciens trong s hnh thnh khi u ngy nay c s dng ph bin chuyn gene thc vt. Agrbacterium gm cc loi A. tumefaciens v A. rhizogenes. A. tumefaciens l vi khun t gram m, hnh que. Vi khun ny c h Rhizobiaceae c kh nng gy u thc vt. N l mt plasmid di vo khong 120-150 kb, c tnh cht gy u nn gi l plasmid Ti. Ti bt ngun t ch u ca tumer inducing. Cn A. rhizogenes l plasmid c kh nng gy bnh r tc (hairy root) v gi l plasmid Ri. Ri bt ngun t ch u ca root inducing tc l sinh r. Kch thc plasmid ny tng t plasmid Ti. i vi cc cy trng, ngi ta c th to ra ging mi cng c nhng c tnh mong mun qua vic chuyn gene. Cc gene c th t mt cy khng c quan h h hng hoc t ng vt, Cn trng, nm men, hoc t cc vi sinh vt. Cc cy c nhn gene bng nhiu cch khc nhau nhng gn y nh h thng vi khun Agrobacterium c c tnh sn c trong t nhin l chuyn c gene ca chng vo cy ci thng qua mt phn T-DNA ca plasmid Ti hoc Ri. Chnh T-DNA ca vi khun
c gn vo b gene ca thc vt s lm xut hin khi u hoc r tc v chng cha oncogene.
Hnh II.2. M hnh plasmid Ti
T-DNA c kch thc 25 kb trong cha gene m ha cho sinh tng hp auxin, cytokinine, opine v cc gene gy khi u (oncogene). Trong plasmid Ti, v tr ca T-DNA c gii hn bng b phi v b tri. Ngoi plasmid Ti cn c cc vng DNA m ha cho vic ti sinh plasmid, cho kh nng ly nhim (vng vir) v chuyn np, cho vic tiu ha opine. Trong cc vng DNA ca plasmid Ti, ngoi T-DNA c nghin cu nhiu hn c l vng DNA ph trch kh nng ly nhim cn gi l vng vir. Sn phm hot ng ca cc gene nm trong vng vir c tc ng kch thch ca cc hp cht phenol tit ra t vt thng l mt loi protein c bit nh vir E2, vir B, vir D, vir D2, vir C1Cc protein ny nhn bit cc vt thng cc cy ch thch hp (hu ht l cy 2 l mm), kch thch sinh sn ra cc on T-DNA bao bc che ch cc on DNA ny v gip chng tip cn vi h gene ca cy ch mt cch an ton. Khi cy nhim bnh do T-DNA np vo trong h gene ca cy ch bt u hot ng v sn sinh ra auxin, cytokinine v opine, ton b sinh trng ca cy b ri lon, cc t bo phn chia v t chc v to ra cc khi u. Opine c vi khun s dng nh mt loi thc n nh gene chuyn ha opine trn plasmid Ti. C ch ly nhim ca A. rhizigenes i vi cy 2 l mm cng tng t nhng trong vng T-DNA ca A. rhizogenes ch c gene sn sinh ra auxin, v th s thay i hnh thi chnh ca thc vt l chng to ra rt nhiu r tc khi b nhim bnh.
Trn thc t bnh cy, Agrobacterium ch gy hi cy 2 l mm. V vy, ngi ta cho rng chng ch c th np T-DNA vo h gene cc cy 2 l mm . Gn y nhiu tc gi chng minh khi nhim vi khun, cc cy 1 l mm cng c th sn xut opine v c th khai thc kh nng bin np gene ca Agrobacterium cy 1 l mm. Nhng nu phn T-DNA b ct th plasmid Ti ny vn gi nguyn kh nng chuyn gene l vo thc vt. Do , plasmid Ti hin nay tr thnh mt cng c sc bn theo c tnh gy u vo cc cy trng. Vic chuyn gene l c tin hnh theo nhng bc sau: * Tch plasmid Ti t vi khun A. tumefaciens * Ct b phn T-DNA ca plasmid i * Chun b gene l cha di truyn mong mun, chng hn gene c t ca Bacillus thuringiensis tiu dit su b hi cy trng. * Chun b gene nh du theo di vic chuyn gene l vo thc vt c thnh cng hay khng? Chng hn, gene m ha cho mt enzyme c phn ng mu m thc vt t c nh gene m ha enzyme glucuronidase gi tt l GUS-A to ra mu xanh da tri c trng vi c cht 5-bromo-4-chloro-3-indolyl, -galactopyranoside c k hiu l Xgluc. i khi ngi ta s dng kh nng nh du ca gene m ha luciferase - mt enzyme ca om m pht sng trong ti cc m c chuyn gene. Cng c khi s dng nhng gene khng khng sinh kanamycine - mt cht c ch sinh trng thc vt. * Gn gene l v gene nh du vo plasmid Ti thay th vo ch TDNA b ct b ri a vo vi khun Agrobacterium. *Chun b t bo vt ch tip nhn vi khun Agrobacterium cha plasmid c gene l nh chun b cc protoplast t cc th m, cc a l. * Nui cy chung vt ch vi vi khun A. tumefaciens cha plasmid Ti gn gene l mt thi gian vi ngy nhit nh sng thch hp. * Loi b vi khun bng dng cc khng sinh c hiu nh carbenicilline. * Chuyn nguyn liu thc vt vo mi trng dinh dng nui cy t bo v m ti sinh c thm nhng cht kch thch sinh trng nh 2,4 D, auxin khc v.v. * Cy ti sinh c nui trong vn m v cui cng trng ra t. * Theo di cc c tnh ca gene l biu hin cy trng. Cn ch : - Cy c bin i di truyn cn phi c trng trong vi v;
- Cy ch ca vi khun Agrobacterium c th lm hn ch s bin i trong nhiu v thu hoch. - Mun chuyn gene thnh cng cn phi phn tch m rng b gene thc vt, bit r t chc phn t ca gene a vo v mi quan h gia n vi t chc cu trc di truyn v iu ha di truyn. i vi s chuyn gene thc vt, virus thc vt cha DNA cng c coi l vector. Loi virus khm thc vt ca hoa sp l (Cauliflower mosaic virus: CaMV) l mt v d hp dn. l mt vector c th dng chuyn gene, v DNA ngoi lai c th chuyn vo thc vt qua s gy nhim l bi virus. Thm vo CaMV c th nhn ln trong bo tng v loi tr kh nng gy tr ngi cho cc chc phn ca t bo ch. S.H. Howell v cng s nm 1987, gi c phn t linker EcoRI gm 8 i base vo vng intergeneic ca virus khm hoa sp l m khng lm suy yu s ly nhim ca chng. Kh nng CaMV nh mt vector chuyn gene c chng minh bi Cronenbor v cng s khi a vo l c ci on operator ca operon lac E. coli v khi DNA ca virus c tch t nhng cy b nhim thy cha on operator lac. CaMV c mt s hn ch: ch gy nhim cho h ci nh bp ci, xi xon, ci Bauxel. Th 2 l ch mt lng nht nh DNA ngoi lai c gi vo (khong 250 i base). Vic dng h vi khun Agrobacterium c hng trm c s cng nghip v phng th nghim trn th gii s dng. Ch Monsanto c hn 45.000 dng thc vt chuyn gene t do c sn xut theo kiu ny. Tuy vy, i vi cc cy la, ng, la m vn phi l cy ch t nhin i vi Agrobacterium. Do , phi a DNA ngoi lai vo protoplast thc vt. Song, kh nng ny lai to c rt thp v mt khc li khng c nguyn vn, phi ph v t bo lai ghp. Do , ngi ta c mt bin php n gin hn l tim DNA ngoi lai trc tip vo t bo. Song cng vp phi mt iu l tim trc tip nh vy kh c hiu qu cao l do: . kim nh d gy v tc; cng vic bun t, t l DNA ngoi lai l t hp vo b gene t bo ch rt thp. Tim khong 10.000 t bo mi hi vng c 1 t bo c gene mi nng cao hiu qu t hp gene. Joh Sanford trng i hc Tng hp Cornell ngh dng bin php bn ph lin tc cc nguyn liu gene vo cc t bo thc vt. Tc gi dng vin n kim loi c ng knh 1-2 c ph DNA ngoi lai. Lm tng nhanh dn tc nhng vin n kim loi ny xuyn vo thnh t bo nguyn vn v phn tn DNA trong bo tng. Nm 1987, h ci tin bng vin n bch kim bn vo t bo thc vt v sau h dng n bng vng v y i bi s bc hi ca git nc.
Vic bn DNA vo t bo tuy ch l giai on u ca vic lm thay i cy trng nhng trc phi kt cu cho c 3 vng quan trng ca gene. l: * Vng promotor vng pht ng biu hin gene. * Vng m ha - to ra nhng protein mong mun thu hoch tt ma mng. * Vng poly A (polyadenine ha) vng tham gia m u sao chp RNA thng tin.
Cng ngh gene cng c kh nng to ra nhng thc n trong sch. Cc gene i vi nhng protein c phm cht dinh dng siu ng c tch ra, sau , gi gene vo cy. Cy ny c th sn xut
ra nhng cht ha hc c bit nh tinh bt, du cng nghip, cc enzyme, cc dc cht . Hn 400 php th ng rung ca nhng cy trng c cng ngh ha M, chu u. Nhng th nghim xc nhn hiu qu kinh t. nhng cy trng c du hiu tt trin khai ra ng rung t nm 1990. Tuy nhin, trong cng ngh gene cng c mt s hn ch nh: ch ci tin mt s im biu hin khng qu 3-5 gene. Mt vi cy trng khng thch ng phng php chuyn gene hin hnh. S tch gene c ch chuyn cng c gp nhng kh khn nht nh. Vic s dng gene trong vic to ging mi nhng cy, hoa, qu,, G. Glili dng vi khun E. coli chit xut ly gene m ha loi enzyme tng hp lysine, sau a vo khoai ty v thuc l lm cho khoai ty lng lysine tng c 5 ln, l 4 ln v thn 3 ln. Ngi ta cng a 9 acid amin khc vo khoai ty lm cho cht lng ca chng tr nn qu gi. M cng va thnh cng nhng qu c chua chn mng mc trn cnh nho. C chua loi ny t thi hng. Ngi ta cng to ra bng c mu sc t nhin bng CNSH. Php mi y, ngi ta cng to cy bp ci to cao hn ngi bng thao tc di truyn. Trong 20-30 nm li y, CNSH ang lm i mi nn nng nghip trn ton th gii nh k thut nui cy m v t bo, phng php chn lc dng soma, k thut dung hp protoplast v c bit s dng cng ngh gene trong ti t hp DNA, ngi ta a vo cy trng nhng nguyn liu di truyn mi, nhng gene c ch lm thay i c bn nhng loi cy trng, to ra mt lot cc ging mi c nng sut cao, phm cht tt, chng chu gii trong iu kin bt li ca mi trng tng khu vc, c nhng ha hn ma mng tt p trong hin ti v khng ngng pht trin trong tng lai khng xa.
nc ta, ng v Nh nc ang quan tm n vn khoa hc v cng ngh, c bit u tin CNSH (nm trong 4 u tin chung: tin hc, CNSH, vt liu mi v hin i ha nn cng nghip). Vi khong 20 triu lao ng nng nghip, c nhim v sn xut ra 20-25 triu tn lng thc hng nm, Vit Nam ng hng th 2, th 3 trong xut khu go th gii, mun lm cho nn nng nghip khng ngng tng ln, vic to ging mi cy trng t ra cho cc nh khoa hc mt nhim v nng n v vinh quang. Di y chng ti xin gii thiu 2 phng php to ging mi hin i chng ta tham kho:
Hnh II.3. Bng phng php trc tip (dng sng bn gene) v phng php gin tip (Agrobacterium) chuyn gene vo t bo thc vt 3. CNSH trong vic bo v cy trng. Hng ngn nm nay, ngi nng dn lun lun thay i cy trng, thay i li canh tc c nhng tin b trong thu hoch. H tuyn chn
nhng ging cy trng ngy cng c nng sut cao phm cht tt hn. Phm cht cng tt th l mn n ngon lnh cho mun vn vi sinh vt, su b. Theo thng k, su b gy thit hi ln cho ma mng khong 2040%. Nng sut ma mng cng cao th thit hi cng ln. Ngi nng dn t xa n nay, qua kinh nghim thc t cng c nhiu bin php phng tr nh xi xo, chm sc bt su b, thay i v trng hp l, trng thm xen k, phun thuc tr su v.v. Ring nc M, mi nm mt i 2,5 t USD v thuc tr su bo v ma mng. Nhng vic s dng thuc tr su bnh, thuc tr c di (herbicide), v d, thuc tr nm ch c phun tr su trn cc vng trng chui ngt (sweet banana) ni ch sn xut khong 10% sn lng chui a vo th trng quc t, 90% sn lng chui cn li l chui bt (platain), mt trong cc ngun lng thc ca nhiu nc nhit i th ch yu trng cc qui m gia nh ri rc. V vy kh s dng thuc tr nm v gi thnh khng kinh t. Hn na thuc tr su bnh li hu qu khn lng, ngi b ng c, thm ch ung th, ng vt c th cht. V vy, cng tc bo v cy trng theo hng CNSH l tt hn c. Ngi ta nui cy nh sinh trng ca khoai ty, du ty, khoai m (Yam) cn gi l khoai n , lm sch virus. Nui cy nh sinh trng cng gip to ra cc ging sn sch bnh. K thut nui cy m in vitro cng gip gii phng cy sa khi bnh virus v Mycoplasma rt nghim trng i vi da, ch l (Date palm), khng bnh hoc t nhim bnh nm hoa t nhn v tnh, c bit l chng chu bnh bayoud do nm (Fusarium oxyporum f-sp-albidinis) gy ra l bnh gy tn tht nghim trng cho ch l. Vin IRAT (Institute de Recherches Agronomiques Tropicales et des Cultures vivrieres: Vin Nghin cu Nng nghip nhit i v Cc cy lng thc) ca Php xy dng c phng php cy tc nhn gy bnh vo cy con ang ti sinh t callus, Vic sng lc cc t bo ca nhng cy con ny c th cho php phn lp c cc dng khng bnh, khng 3 loi bnh quan trng: sinut, ieafscala v bnh r st. Tng t nh vy, ngi ta c th tch c cc dng t bo khng mn, khng phn v c t ca nm (Helminthosporium sacchari). Cc kt qu nghin cu ca CENA (Centre de Nuclear Energy in Agriculture) v CEBTEC (Diviso de Biotechnology de Plantas et Centro de Biotechnologia Agricola) Brazil cho thy dung hp protoplast cy Ma c th gip chuyn gene khng thuc tr su c t dng ma ny qua dng ma khc. T nm 1985, phng Hin vi in t ca CENA bt u chng trnh nghin cu cy chuyn gene khng bnh khun v virus vo t bo cy trng, sau tch protoplast v dung hp chng trong in trng.
Mexico, cc tc gi Merino v Madrigal trng i hc Chapingo cng tc vi cc nh nghin cu ca i hc Purduc (M) th tch cc dng bin d khng 2 ni r st ang gy hi chu M Latinh trn i tng cy c ph. Dung hp protoplast gip cho c ph khng bnh v ngi ta cng dng c t do nm ny tit ra da vo mi trng nui cy nhm chn cc dng t bo khng c t. Ngy nay cng ngh gene c dng to cc ging cy chng cc bnh virus, khng cc cn trng c hi, to ging mi chng c di, chng h hi ma v cy trng. Roger Beachy trng i hc Tng hp Washington v Stephen Roger Monsanto p dng cng ngh gene cu trc nn mt vector gene da vo cy thuc l v cy c chua mt protein ph mt ngoi ca virus khm thuc l (tobacco mosaic virus: TMV). Nhng cy c i khng rt mnh m vi nhim trng v nh vy xc nh gi thuyt ban u ca Beachy l ng - mt thnh phn n gin ca virus c th s dng bo v c s tn cng ca virus. Ngy nay, s biu hin mt protein ph mt ngoi ca virus no c th s dng chng virus thc vt tr thnh mt c ch chung bo v cy trng khi bnh virus. Trong vng 30 nm qua, ngi nng dn v cc nh lm vn tin tng v vi khun Bacillus thuringiensis (Bt) v chng sn xut ra protein chng Cn trng. Vi khun ny lm cht hng lot su t, Su rm vng Thuringi. Cc ch phm Bt c hiu cao vi cc cn trng thuc loi bm. C ch hot ng phn t Bt nh sau: protein Bt lin kt vi nhng proceptor c hiu c khu tr trn mng rut ca nhng cn trng gy hi. S lin kt gy cn tr cho dng vn chuyn ion vo trong t bo biu m rut v nh vy lm mt kh nng n ung dinh dng ca cn trng. Nhng thuc tr su thin nhin ny khng gy c hi cho ngi v ng vt, thm ch ngay c cc loi cn trng khc. Mt khc, tnh cht c ch ca thuc tr su Bt thng b hn ch v d dng b ra sch khi cy, nhng hiu qu ca chng trn ng rung thng c ko di. Gia nhng nm 1980, cng ngh gene ca mt vi hng ln nh hng H thng gene thc vt Belgium, hng Di truyn Nng nghip Middleton v.v. thnh cng trong vic tch ri cc gene vi khun m ha cc protein thuc tr su. H dng sng c Agrobacterium tumefaciens c gn nhng gene tch ni trn bn vo khoai ty, v chua v bng. Lc u nhng gene ny biu hin ngho nn. Nhng protein Bt c cy trng sn xut ra ch git c nhng cn trng phng th nghim nhy cm.
Cc nh khoa hc Monsanto nh David Fichhoff v Frdrick Perlak ci tin. H xem xt k li gene vi khun nguyn thy bt chc cht ch hn on DNA thc vt. Mt s ci tin nh lm tng ln s kim tra cn trng mt cch mnh m. Hai nm th nghim trn ng rung xc nh rng s c mt ca cc gene Bt trong cy bng kim tra mt cch c hiu qu tt c nhng h hi ca i b phn cc loi bm su bao gm cc mng trng ca chng. Nhng cy bng c thc hin cng ngh gene lm gim hn vic s dng thuc tr su t 40 n 60%. Cc nh khoa hc sng lc mt cch m rng i vi nhng dng Bacillus thuringiensis t nhin. Nhng dng c hiu qu trn cc cn trng khc hn l loi bm. H chn c mt dng c mt gene c hiu qu chng li loi gin cnh cng ph hoi khoai ty: Colorado. Ma xun nm 1991, Russet Burbank ghp gene kim tra loi cnh cng ny trn khoai ty v th mt vi ni Maine v Oregon. Khoai ty min dch c loi cnh cng ny. Cc nh khoa hc t hp Mycogene San Diego pht hin mt gene Bt chng Giun trn k sinh thc vt v mt gene chng Mui. Hin nay ang c gng sn xut protein tr Mui (mosquitocidal protein) To s dng vo bin php chng st rt. Tnh c hiu ca protein Bt v s khu tr ca n trong cc m ni trn l protein ny ch chng li bn Cn trng tn cng. Protein khu tr ni bo ny chc chn khng b ra sch v thuc loi thuc tr su an ton nht. Bn cnh vic khng virus, Cn trng gy hi cc cy trng cn phi chng i cc loi c di. C di lm gim sn lng ma mng c khi ti 70%. Trc y ngi ta phi hp thuc dit c vi vic lm c hn ch c di. V trong thuc dit c c ph hn ch trong hot ng, n ch gy nh hng rt nh i vi c di. Cng ngh gene c th dng kim tra mt cch hu hiu c di. Vn l ch to ra cho cy trng c th sn xut thuc tr c di vi ph rng, n gin v an ton cho mi trng. Nu s dng c cng ngh vo cho cy trng s lm cho nng nghip gim mt lng ln thuc tr c di. C 3 hng cng ngh to ra thuc tr c di: * Cc nh nghin cu Monsanto, Calgene, California dng thuc tr c di tn l Roundup. Roundup l thnh phn ph rng tr c di l to v mc dy. Roundup c ch hot ng enzyme tng hp EPSP enzyme ny tham gia vo vic to acid amin thm cn cho s sinh trng
ca c di. Roundup khng lm nh hng ti ng vt (ng vt khng tin hnh to acid amin thm). Hn na, Roundup li thai ha nhanh chng trong mi trng tr thnh v hi. Nm 1983, Luca Comai v David Stalker Calgene, Roger v Ganesh Kishore Monsanto tch c gene ca enzyme tng hp EPSP t vi khun v cy trng. Sau cc nh khoa hc to nhng gene sn xut ra lng protein ln hn ln trong cy trng ri a vo c chua, u, bng, ci du, v mt s cy khc, cc cy trng c th tr c c di mt cch c hiu qu. * Cc nh nghin cu Du Pont dng k thut nghin cu tng t tin hnh cng ngh gene ha nhng cy trng c th khng dng thuc dit c sulffonylurea. * Cc nh khoa hc H thng di truyn thc vt v hng Hoechst - c c hng tm kim khc loi tr thuc dit c. T vi khun Streptomyces hygroscopicus tch ra mt gene enzyme c ch cht dit c c gi l Basta. Basta nh hng ti vic tng hp glutamine ca enzyme trong c di v lm cho c di khng ln ln c. Nhng trc khi gy hi, c th xy ra nhng cy trng c gene lm c ch Basta. Trong bo v qu t b h hi, cng ngh gene cng ang pht huy tc dng. Ngi ta xc nh v tch mt s gene ng vai tr trong sinh tng hp ethylene - mt phn t lin quan n s chn ca hoa qu. S chn ko chm li kp thu hoch v to mi tt hn lm phm cht tng ln. C 2 phng php di truyn tng ngun thu hoch qu. Mt l gi on dch m antisen ca gene chn lin kt vi RNA thng tin lm tt gene. Athanosios theologis California v Don Grierson trng i hc Tng hp Nottingham chng minh rng, cc qu C chua c nhng gene antisen chng s lm mm. Th hai l a gene vo to enzyme lm thai ha thnh phn tin cht hnh thnh ethylene, nh vy s lm chm s chn v h hng (cng trnh ca Kishora v Harry Klee Monsanto). Gn y cc nh khoa hc M cy vo cy 1 gene vi khun sn sinh ra 1 cht chitinase tiu dit cc t bo ca nm (cy vo c chua, khoai ty, rau dip v cc ging cy tng t nhng cha lm c i vi la, la m, ng v cc cy c ht khc). Ngoi CNSH ra, hin nay ngi ta cn dng 1 s bin php sinh hc thay th thuc tr su ha hc nh; nui ong mt dit Su ay, dng ht cy su u dit gn 2000 loi su hi (trong su u c azadirachitine c tc dng dit cn trng mnh). n xy dng nh
my cng sut 20 tn ht su u/ngy. Php, ngi ta dng b ra dit rp. Mt con 1 ngy n khong 100 con rp hi cy n qu, nn ngi ta gi nhng con b ra dn vn. M, ngi ta c tnh c di Leafy Spurge 7 nm ti s mc dy 2 triu ha t trng Bc M, Canada, v s lm tn tht hn 100 triu USD. Ngi ta dng b cnh cng tiu dit c di. c, hng ngn ha rng Si b mt loi nhn tn ph trong ma m chi. Ngi ta phun Bacillus thuringiensis ln ngn cy. Khi cn trng n vo cng vi chi non, l non th lm cho chng khng mun n na v sau chng b thng bng, ln ra cht. M, ngi ta dng loi virus polyeder sn ui v git cht cc loi su v cn trng c hi, nht l loi nhn c. Vit Nam ta th nghim nui su xanh cho trn thc n c virus. Sau 6-8 ngy mi virus thnh 10.000 virus khc, tc l chim khong 30% trng lng kh ca con su. Khi su cht, gp ring ra, nghin nt ly nc dch. Dch ny li em phun dit su xanh phi hi cy trng. Nm Boveroa cng c dng tr su rm thng. Nm Metthirum cng ang c nghin cu tr ry nu hi la. Virus NPV - virus a nhn (nuclear polyhedrosis virus) v GV (graluclar virus) ang c s dng vo vic dit su bng. Hin nay ngi ta cng dng pheronom - cht ta ra mi hng dn d cn trng tp trung vo mt ch m tiu dit. 4. CNSH trong vic sn xut phn bn Nitrogen l mt trong nhng nguyn t c bn ca s sng, bi v nitrogene chim khong 16% trong protein. Nitrogen li l thnh phn quan trng ca acid nucleic m protein v acid nucleic l thnh phn quan trng ca s sng. Ta bit rng, trong khng kh c 80-150 tn N2/ha tc l gp vn ln so vi nitrogene trong t. Nhng N2 dng phn t rt bn chc dng kh tr cy khng ly trc tip c. V vy, con ngi phi to ra nhng nh my khng l v rt tn km bin N2 thnh dng phn m urea, m nitrate, nitrite bn cho cy. S lng m trong nh my khng cho nhu cu dinh dng ca cy (ch mi cung cp 2-3% lng m cho cy). Trong t c mt s vi sinh vt t c kh nng c nh N2 trong khng kh, vi khun cng sinh vi cy b u, cc to lam sng t do hoc cng sinh vi bo du sn xut ra phn bn cho cy trng.
Hin nay, ngi ta to ra nhng ging cy trng c nng sut cao thng nhu cu i hi v phn cng cao. V vy m t khng p ng c nhu cu cho cy. Cc nh khoa hc ngy nay pht hin nhng b quyt ca vi sinh vt c nh N2, tng lng tiu tn rt t m hiu qu ca s c nh cao dng lm ngun phn bn bng con ng CNSH. 4.1. Nhu cu phn bn i vi cy trng n nm 1974, ngi ta sn xut c 4.106 tn phn m tr gi 8 t dollars a vo s dng cho cy trng. Trong khi 25 nm v trc hng nm mi ch c 3,5.106 tn. n cui th k XX, do nhu cu v lng thc ngy cng tng, con ngi cn phi sn xut 200.106 tn phn m. R rng nng lng dng tiu th cho cc nh my phn m sn xut s lng phn nh trn th rt kh khn. Do , ngi ta phi dng nhiu bin php k thut khc nhau: phng php cng nghip, phng php sinh hc, phng php di truyn. 4.2. Cc phng php k thut khc nhau trong vic c nh N2 * Phng php c nh N2 cng nghip bng qui trnh HaberBosch. Bng phng php ny ngi ta xy dng cc nh my kh tn km: cn nhit cao (500oC), p sut ln (hng ngn atm) cng nh s c mt ca cht xc tc bin kh N2 + 3H2 2NH3. Hin nay c nh my ln sn xut 1.000 tn/ngy, nn th gii mi nm sn xut c 4.106 tn phn m. Song hn ch ln nht ca phng php ny l tin vn u t qu ln. Tuy vy, do nhu cu ca phn bn m ngi ta xy dng khong 400-500 nh my. * Phng php c nh N2 sinh hc bng cc vi khun. - Vi khun nt sn cy b u. Ngi ta pht hin vi khun hnh que c chiu di khong 0,5, c tim mao, n nu trong cc u sn ca r cy b u, l Rhizobium. Nt sn l nh my phn m ngm di t ca cy b u. Trong vi khun ny c mt gene iu khin vic tng hp enzyme nitrogenase v hydrogenase cng vi sc t mu hng bin N2 phn t thnh NH3. Nh h enzyme hot ha ny m Rhizobium ch cn mt nng lng nh (3-5 kcalo/M) v cng khng cn p sut cao cng c th d dng b gy cc ni lin kt nitrogen trong phn t (NN) d dng hnh thnh NH3. (Mun b gy ni ba th nht cn 215kcalo/M, b gy ni ba th hai cn 127,65 kcalo/M v ni ba cui cng ch cn 38 kcalo/M). Vi khun nt sn cy b u c th ng ha 100-250 kg N2/ha.nm, c Luzern: 300, c Stylo: 150-200, u: 80-120, vi khun sng t do trong t: 25-40 kgN2/ha.nm. Vi sinh vt trn v qu t c kh nng ng ha 100 triu tn/nm.
Ngy nay, ngi ta sn xut phn bn nt sn Rhizobium bng cch phn lp cc vi khun nt sn, tin hnh nui cy nt sn ny ni ln men ln vi cc mi trng dinh dng thch hp (ngun hydrocarbon v protein). Khi dung dch nui cy t c mt lng ln vi khun em ra trn ln vi than bn kh c nghin cng vi r ng, ng ti nh h ming ti t 3-5 ngy nhit 20oC ri dn ti, bo qun trong t lnh v chuyn n ni tiu dng . l loi phn sinh hc nitragin. - Vi khun lam (to lam) c nh N2 T nm 1910, Bottomley cho rng trong ti l bo du ngoi to lam Anabaena cn c cc loi vi khun khc nh Pseudomonas radicicola v cc loi Azotobacter. To lam cung cp cho vi khun cc sn phm quang hp cn vi khun cung cp nitrogen c nh c cho to lam. Vit Nam, L Vn Cn v mt s ngi khc (1954) nui 1 gam bo du trong dung dch khng cha nitrogene, sau 20 ngy thu c 42,6 gam. Lng nitrogen trong bo du t 2 mg tng ln 75,4 mg. Trong 2 thng lng nitrogen m bo du c nh c 136 kg/1mu Bc B. V vy, bo du l cy phn xanh c gi tr cho ngh nng. - To xanh lc dng sn phm quang hp ca mnh lm ngun nng lng ng ha N2 ca kh quyn. V vy, to xanh lc trn cc rung la ang l vn thi s c nhiu ngha trong vic tng lng m cho cy. - Vi khun nitrogen sng t do trong t . Chlostridium pesteurianum l loi vi khun ym kh. Qu trnh c nh N2 thng din ra nh sau: T qu trnh ln men butyric: C6H12O6 C3H7COOH + 2CO2 +4H+ Hydrogen ca qu trnh ny Chlostridium ly kt hp vi N2: 2N2 + 6H+ 2NH3 nh Chlostridium c hai tiu phn hot ha hydrogen v nitrogen (hydrogenease v nitrogenease) khi s dng 1 gam ng th Chlostridium pasteurianum ng ha c 2-3 mg N2. . Azotobacter l loi vi khun hiu kh nh c tnh oxyha hiu kh trong qu trnh trao i cht. Cho nn hiu qu c nh N2 ca chng ln hn. C 1 gam ng c th c nh c 15 mg N 2 thm ch chng t ti 30 mg. Bn cnh , cc vi khun ny cn cung cp mt lng ln cht iu ha sinh trng nh auxin, gibberelline. Do tng c nng sut cy trng trong c la. 4.3. Cng ngh gene trong sn xut phn m
Cng ngh gene trong sn xut phn m cho cy trng l mt trong nhng vn sinh vt hc hin i nhm s dng kho tng N 2 phn t qu gi v v tn ca kh tri m trc y thin nhin ch dnh cho Rhizobium, Chlostridium, Azotobacter, Azospirillum (gn y ngi ta pht hin ra t bo r la m, la nc, ng, c mt loi vi khun sng cng sinh gi l Azospirillum trong A. brasilence la m, A, proferum ng) hay nhng x khun ca c ph, phi lao, hay vi khun mt s cy tng bch thuc ru, thuc tu, v.v. Nhng thnh tu gn y trong sinh hc phn t nh vic pht hin enzyme phin m ngc dng (revertranscriptase) m ra s tng hp gene nhn to ca Temine, Baltimore, vic pht hin cc enzyme ct hn ch (restrictases) cc on gene trong b gene t bo lai ghp cc ti t hp gene mt cch d dng ca Arber, Nathan, Smith to iu kin cho cc nh bc hc M c th bin c cc trc khun ng rut E, coli vn khng c kh nng c nh N2 thnh mt chng sc tiu th c N2. Enzyme nitrogenease v s hot ng c nh N2 ca n. Nm 1960, nitrogenase c tch t Chlostridium pasteurianum. Lng enzyme ny cha ti 5% protein ca vi khun. Khi cho thm mt lng ln pyruvate vo mi trng nitrogenase hai thnh phn sau: 1 thnh phn cha Mo v Fe gi l protein Mo-Fe. Thnh phn th 2 cha Fe v gi l protein-Fe. Protein Mo-Fe c lm kt tinh v c cu to t 4 n v con sp xp thnh mt hnh hp nh quan st trong cc nh hin vi in t. Protein ny cha 2 nguyn t Mo v 24-32 nguyn t st v lu hunh. M=220.000dalton. Protein st c cu to t 2 n v con ging ht nhau, cha 4 nguyn t st v 4 nguyn t lu hunh M=60.000 dalton. Ph cng hng spin in t protein Mo-Fe l duy nht i vi cc cng hng cc gi tr ca g bng 2,01; 3,65; 4,30 thuc v s mt nguyn t st. Ph ny c chng minh l c ch i vi cc kho st v c ch v cc kho st v sinh l. Cn ph cng hng spin in t ca protein-Fe khng phi duy nht m tng t nh ph quan st c i vi ferredoxin. C 2 thnh phn ni trn u c vai tr c bn i vi hot ng ca nitrogenase theo t l 1 hoc 2 protein-Fe i vi protein Mo-Fe. Ngi ta thy protein Mo-Fe c tch ra khi c th ny c th ti hp vi protein-Fe c phn lp mt c th khc to ra 1 enzyme nitrogenase c chc nng. iu chng minh thm cho s ging nhau ca cc nitrogenase t cc ngun khc nhau. Ammoniac l mt sn
phm ca s c nh N2 sinh hc. NH3 khng phi l mt cht c ch cc phn ng. ATP cng nh 1 cht kh thch hp, u c vai tr c bn i vi s hot ng ca nitrogenase. di chuyn c 1 electron, enzyme nitrogenase cn 4 phn t ATP. Ferredoxin hoc flavodoxin c th oxyha-kh gn in cc hydrogen l nhng tc nhn sinh l chuyn electron duy nht bit c lin kt vi nitrogenase. Pyruvate l cht cho electron t nhin vi sinh vt nh Chlostridium , cn Rhizobium hnh nh l phosphate dinucleotid adenine nicotinamid kh (NADP.H2). Nitrogenase c tnh linh hot tuyt diu v kh nng kh hng lot nhng cht khc nhau. Mt enzyme khc cng c tnh cht linh hot l ribulosodiphosphatecarboxylase enzyme c nh CO2 v O2. C th xem nitrogenase nh mt reductase i vi H3O+ v i vi cc lin kt ba do cc chc NN, NO, NC, v CC biu th bng N 2, N3, N2O, RCN, RNC, v RCCH cho sn phm biu din cc phn ng thm 2, 4, 6, 8, 10, 12 v 14 in t. ng lc ca phn ng nitrogenase cho bit phc hp ATP v Mg lin kt vi protein-Fe ch khng phi protein Mo-Fe. Nhng thay i trong ph cng hng spin in t ca protein-Fe v protein Mo-Fe dn n kt qu l protein Mo-Fe b kh bi proteinFe vi s tham gia ca ATP, Mg. Nh vy, nitrogenase c biu th nh mt phc hp ca protein-Fe v protein Mo-Fe vi Mg-ATP bao hm trong qu trnh chuyn in t t protein-Fe sang protein Mo-Fe. * Chuyn gene sn xut nitrogenase (gene nif) t vi khun c nh N2 sang E. coli bng con ng k thut gene. Cch y hn 10 nm, cc nh bc hc M tch mt on gene iu khin vic sn xut nitrogenase ra khi vi khun Klabsiella pneumoniae ri ghp on gene ny vo vi khun E. coli mt cch an ton. Khi vo E. coli, gene ny bt tay vo vic sn xut nitrogenase. Enzyme ny gip trc khun th E. coli d dng tip thu nitrogen phn t m bnh thng E. coli khng thc hin c. * K thut gene trong chuyn operon nitrogen vo cht nguyn sinh ca cy trng mong mun. Ngy nay ngi ta mong mun chuyn gene nif t vi khun c nh N2 vo thc vt thng ng nh la , khoai, cy n qu v.v. lm cho chng c kh nng t tc v phn m. Hi vng rng, trong thi gian n, bng k thut gene, ngi ta chuyn cc plasmid cha gene nif vo cc cy trng v s chm dt nn i m trin min ca cy trng. Lc by gi chc chn s to ra mt bc ngot v i i vi s pht trin nng nghip trn ton th gii.
5. CNSH trong vic nghin cu cht HST lm tng nng sut cy trng. Ta bit, cc t bo c th (soma) thc vt c tim nng trong mt iu kin no hon thnh mt cy hon chnh. K thut ny cho php ti sinh t nhng t bo c th b bin tnh hay t bo sinh sn n bi, khai thc s khc nhau ca dng t bo c th ca cc m thc vt v pht trin cc k thut di truyn ging hiu lc lm tng tc truyn ging ca cc cy trng mi v khng c virus. iu ha s sinh trng v pht trin ca thc vt bao gm ti sinh cc thc vt t nhng t bo v m tch ri u di s kim sot ca hormone gi l phytohormone. Phytohormone gm nhiu cht nh: auxin, abscisin, ethylene, gibberelline, cytokinine. Nhng 2 loi hormone quan trng hn l auxin v cytokinine quyt nh s kch thch phn chia v bit ha t bo ca cc m c nui cy in vitro. Vic nui trng bng m lu di ca mt s loi thc vt c tin hnh nh cng thm auxin vo mi trng. Sau , ngi ta pht hin ra cytokinine (xem thm Gio trnh l thuyt v Sinh l thc vt ca Trng Vn Lung, 1991, trang 174184 v nhiu ti liu khc v cht iu ha sinh trng thc vt). Cytokinine t nhin c pht hin u tin l zeatin v tch ra trong ni nh (endosperm) ca ng non. Ging nh cc cytokinine t nhin khc, zeatin l adenine c thay th N+6, vng adenine nguyn vn mang chui bn N+6 vi lin kt i v tr 2,3 v nhm 4 methyl c hydroxyl ha. Hu ht cc nghin cu xc nh mi lin quan gia hot ng sinh hc vi cu trc ha hc ca cytokinine u th nghim trn cc callus ca thuc l. Kh nng cytokinine kch thch s ln ln ca m thuc l c b sung auxin. Cc tc ng sinh hc ca cytokinine - Kch thch s sinh trng ca t bo thc vt cng vi vic thm auxin. - T l auxin/cytokinine cao trong mi trng nui cy th cm ng sinh r, c mnh; ngc li th sinh n, cnh non tng. - S ln ln ca chi bn tng, chi nh th gim, lm d dng cho vic to hnh nhng cnh c kh nng sinh sn. - Tng s khng ca cy ti cc tress khc nhau nh mn, m, nhit cao. - iu ha sinh trng ca cy di nhng iu kin hn hn, lm tr hon s gi yu ca cy v cc phn cn li ca cy b cht. - Kch thch dip lc pht trin.
- Trong mt s trng hp no , cytokinine iu ha s biu hin ca nhng gene c hiu ca mt s loi thc vt. H2 HN - C O N N NH N Kinitine HN-CH2 N N NH N N+6 benzyladenine Thydiazuron (6-benzylaminopurine) (B4) Cc cytokinine Cytokinine xy ra trong thc vt nh l nhng thnh phn t do v nh l nhng cht c cu thnh t vi loi RNA vn chuyn (tRNA1). Cc cytokinine t do trong cc m cy trng b bin tnh tN 6-(2isopentenyl)adenosine 5- monophosphate (t6AMP) chng bt ngun t hai cht tin thn l 5-AMP v A2-isopentenyl pyrophosphate. Enzyme xc tc ca chng l 2-isopentenyl pyrophosphate: 5-AMPisopentenyl transferase (gi tt l ipt hay cytokinine synthetase). Enzyme ny c pht hin trong ch phm v bo ca Diotystelium discoideum. Hot ng ca enzyme cytokinine synthetase ny cng tm thy trong callus thuc l (vic tch chit cng kh khn). Enzyme cytokinine synthetase cng c pht hin trong nhng t bo thc vt b bin tnh bi Agrobacterium tumefaciens. Tnh c ca A, tumefaciens c lin kt vi plasmid cm ng ung th mang nhng gene enzyme sinh tng hp auxin (gene 1 v 2) v cytokinine (gene ipt hoc gene 4). Nhng gene ny c mt trong on N N -CH H -C N O C H N N Trans-zeatine N NH H CH2OH C=C HN-CH2 CH2 N
plasmid Ti ring bit (T-DNA) n c th truyn t vi khun ti b gene ca t bo thc vt ch. S biu hin ca n l sn xut qu nhiu auxin v cytokinine dn ti kt qu c phenotype kiu ung th. Thm vo gene ipt, mt s dng ca A. tumefaciens cng mang gene m ha isopentenyltransferase (tZs), n khu tr bn ngoi vng T-DNA, st vi vng DNA c lin kt vi chc phn c. Gene tZs c biu hin trong vi khun v khng c chuyn ti b gene vt ch. S biu hin ca n c cm ng bi nhng phenolic ca thc vt c to ra sau khi cy b thng. Vai tr ca gene ny c th l cm ng s phn chia t bo dn n hnh thnh t bo rt nhy cm i vi vic nhim trng. C gene ipt v plasmid Ti v tZs c a vo E. coli v nhng nghin cu gii m tng on ch ra s ng nht ca chng. Ngoi ngun gc vi khun ca n ra, ipt l gene sinh tng hp hormone thc vt u tin gene ny c thun ha v c biu hin in vitro. Ngi ta bit tRNA nh l ngun cytokinine t do. Cc cytokinine c mt trong nhiu cy k c thc vt bc cao v c cu thnh t mt vi loi tRNA, nhiu ti liu cho rng c ti 50% cytokinine t do c dn ra t tRNA. Song thc vt tin ha cao hn c c ch lm ngn cn cytokinine c dn ra t tRNA v c c ch lin quan ti s iu ha hormone. Cis-zeatin l thnh phn cytokinine tri ca tRNA thc vt. Ciszeatin biu hin hot ng cytokinine thp hn nhiu so vi trans-zeatin. Song cis-zeatin c th c bin ha isomer trans bi hot ng ca enzyme cis:trans.isomerase n c tm thy trong phi ca Phaseolus vulgaris. Enzyme ny c th tham gia kim sot s phn phi tRNA zeatin bin thnh cytokinine t do. Nhng cc m ln ln trong ti nh r th n khng hot ng, m r li l ngun chnh ca cc cytokinine thc vt. S chuyn ha cc cytokinine bao gm : - Bin ha cc base, cc nucleoside v cc nucleotid. - N-glucosyl ha v s kt hp alanine ca vng purine. - O-glucosyl ha v acetyl ha ca chui bn. - S kh chui bn. - Lm gy chui bn. S kt hp ca chng c tc dng kh c hay v hot nhng cht hot ng sinh l trong thc vt v ng vt. Chng hnh thnh cytokinineN-glucoside v 9-alanine kt hp vo zeatin v dihydrozelatine. Cc cytokinine-N-glucoside c th dng nh ngun ca cc cytokinine t do trong t bo thc vt b bin tnh bi T-DNA ca plasmid cm ng sinh r (Ri) ca Agrobacterium rhizogenes. Vi khun ny gy bnh r tc trong
cc cy c kh nng kim sot nng cytokinine v auxin ca cc t bo thc vt ch khc nhau. Plasmid Ri ca n cha cc gene cm ng sinh r (rol): rol C v rol B lm m ha cc gene -glucosidase c hiu xc tc s gii phng cytokinine v auxin. Nhng gene hydrolase ny hoc l vng mt hoc l b c ch trong cc t bo thc vt bnh thng. y cng l mt li kh y tim nng cho s iu ha nng cytokinine v auxin ca cc loi thc vt, trong N-glucosyl ha l phng php ch yu lm v hot cytokinine t do. Ngc li cc O-glucoside gm cc lin kt glucosyl, xytosyl v ribosyl ca cytokinine th khng vi s oxyha sinh hc v biu hin hot ng cao trong cc th nghim sinh hc i vi cytokinine. Cc Oglucoside c th lm c ch trc tip hot ng sinh hc. Ngi ta tch c enzyme transferase 0-glucosyl zeatin t phi ca cc loi Phaseolus khc nhau. S to thnh cc khng th n dng i vi cc enzyme c th lm thun ha c nhng gene thch hp. Nhng gene l nhng li kh c ch cho vic nghin cu chuyn ha zeatin (enzyme c hiu zeatin) chng c th s dng sinh cc khng th nhn din cc v tr lin kt zeatin - tc l nhn din cc receptor (c quan nhn cm) ca cytokinine. Enzyme ch yu lm c ch cytokinine t do trong t bo thc vt l enzyme oxydase cytokinine. Enzyme ny xc tc c hiu lm gy chui N6. Nhng c cht t nhin l cytokinine mang lin kt i 2 trong chui bn (c ngha l i6Ade v nhng riboside ca chng, cc kt hp Nglucoside v N-alanyl. S vng mt ca chui i 2 (cytokinine type dihydro-zeatin) hoc s c mt ca cc thnh phn c chui bn nh Oglucoside zeatin v N6-benzyladenine lm cho cytokinine khng vi s thoi ha ca n. Ngi ta chng minh rng, khi cung cp cytokinine ngoi vo th tng hot ng ca enzyme oxydase ca cytokinine. l mt trong nhng l do lm yu cytokinine ngoi lai kch thch s phn chia t bo v s ny chi trong mt s nui cy m thc vt. Nu s sinh tng hp cytokinine trong cc m thc vt khng b bin tnh c xc tc bi ch 1 enzyme n gin th nng cytokinine ca cc t bo thc vt s kim sot bi 2 enzyme: synthetase cytokinine v oxydase cytokinine. Thng thng, cytokinine c a t ngoi vo l lm tng cnh mt s cy cnh. a cytokinine vo cc cy ng cc sau khi ra hoa
s lm tng s ra ht v sinh ht ca la m, la mch, la nc, yn mch v ng. c bit tc dng su sc i vi nhng cy ln ln khi thiu NO3Cytokinine tng hp rt t v c hi cho sc khe con ngi cho nn ngi ta sn xut cytokinine bng cng ngh gene. S chuyn gene ny s tng hp ra cytokinine t nhin trong cc thi im thch hp cho s pht trin ca cy. Vic a gene ipt vo nhm lm chm s gi ho ca cc l ca cc cy ng cc l nhng nghin cu y l th i vi tc dng ca cytokinine. Cytokinine c tng ln khi gene ipt c hp nht vi promotor cu to to ra nhng hiu qu to ln trn cc phenotype thc vt. Vic lm gim nng cytokinine lin quan vi nng auxin trong t bo thc vt c tm quan trng trong thc tin. Chng hn tch nhng phong b sinh l, ngn cn s sinh r v lm gim s to cnh ca mt s cy hoa v cy cnh trang tr bng k thut antisense tc l a nhng on antisense ca gene ipt vo hay nhng gene m ha enzyme oxydase cytokinine, cng nh gene m ha enzyme transferase N-glucosyl c hiu cytokinine bng nhng promotor thch hp. Nng cytokinine m thuc l b gim bi tng nng auxin. a auxin tng hp vo dng thuc l, m s lm tng s thai ha (H 3) zeatin in vitro. Nng cytokinine b kim sot bi s c mt ca gene tng hp auxin. Ni tm li, t l cytokinine/auxin trong cc t bo thc vt, s a gene ipt vo b gene thc vt qua con ng cng ngh gene lm bin i su sc hnh thi cy trng. S tch nhng gene iu ha ca cytokinine, s b gy chui bn lm v hot v d tr li lm sng t vai tr ca cht HST trong c cytokinine. Vic thun ha cc gene m ha cc receptor ca cytokinine l mt trong cc con ng nghin cu kim tra hot ng ca cytokinine v kim tra hiu qu ca chng trong qu trnh tc ng ti nng sut cy trng. 6. Cng ngh sinh hc trong vic to ging vt nui cho nng sut cao 6.1. Cng ngh c truyn trong vic to ging vt nui T thi xa xa con ngi bit thun ha th hoang, chn gi li nhng con tt p ng c nhu cu th hiu ca mnh. Con ngi chn lc ngay t con vt gc theo nhng c tnh di truyn tt tip tc gi gn v khai thc. Nhng con c v con ci tt nht c gi li lm ging. Chng khng nhng c chn theo dng hnh lin quan n kiu di truyn m cn tc ng mnh v mt dinh dng vo qu trnh sinh sn. Hiu qu chn lc tch ly qua thi gian di to nn kiu di truyn phong
ph t phi thai m khi trng thnh, vt nui s biu hin bng kh nng cho tht, sa, trng. Tiu chun chn ging v v khu phn dinh dng lun tc ng mnh n sinh l, trao i cht ca con vt to nn s a dng, phong ph cho kh nng sn xut v sinh sn ca con vt. Con ngi cn bit vn dng nhng t bin c li nng cao sc sn xut ca con vt nh B tht khng sng, cu ly lng ngn chn, g tht tri lng c Khi chn lc c th, con ngi cn chn phi tc l tm cch phi hp cc tnh trng tt ca i b m truyn cho i con. T m c cc phng php to ging vt nui mi, dng mi, cc t hp lai mi nng cao nng sut v tng thm a dng sinh vt cho qun th. Trong lch s chn nui, lai to ging c s dng to ra nhiu ging vt nui qu gi, cung cp vt liu di truyn a dng chn lc v cng c cc t hp gene qu ph hp vi nhng mc tiu c th. 6.1.1. Nguyn tc chung ca vic to ging mi - Trc ht phi c vt liu khi u thun chng v tnh nng sn xut. Ngi ta thng dng nhng c th thuc nhng dng cao sn ca nhng ging thun. Khi phi dng nhng ging a phng hoc nhng ging nguyn thy lm vt liu khi u th ging phi c mt s lng tng i nhiu v phn b trn mt a bn nht nh. - Tin hnh lai to. Sau khi thy cc ch tiu mong mun khng chch hng th s dng cn huyt. Mc v bao nhiu i phi cn huyt ty thuc vo tnh hnh t c cc tiu chun ra mc no trn con vt v tu theo ging em dng c thun chun cao hay thp. - Vic chn lc phi tin hnh rt kht khe, khng nhng ch da vo cc tnh trng hnh thi m c bit phi ch trng n tnh nng sn xut thng qua cc phng php di truyn. - Khi t c nhm ht nhn hay mt dng ca ging mi th vic chn lc, nui dng tip theo mt chng trnh c nh hng v vic p dng cn huyt sau phi thn trng. Lc u cn huyt l cng c cc tnh trng t c, sau l cng c thm cc tnh trng mong mun, ng thi loi thi cc gene ln xc nh cc tnh trng xu (cn gi l lm trong sch dng thun). Song lc ny khng cn thn s dn n suy thoi. - Tip tc to dng v nhn ging theo dng. 6.1.2. Cc phng php lai ging nhm mc ch to ging mi Lai ci tin (topcrossing) v lai ci to (grading up) Hai phng php ny ni chung l rt ging nhau. Lai ci tin
Lai ci tin l phng php lai trong dng con c ca ging i ci tin cho giao phi vi con ci ca ging b ci tin. Sau dng con c hoc con ci ca ging b ci tin phi hp vi con ci hoc con c ca cc i lai (con c ca ging i ci tin ch dng mt ln). Khi t yu cu th c nh (t giao). Kt qu l s c con lai mang ch yu l h gene ca ging b ci tin v ch c mt t l nh ca h gene ca ging i ci tin. Lai ci tin thng p dng trong trng hp khi c mt ging m tnh nng sn xut ca n tng i tt (nh lng sa nhiu) nhng vn cn nhc im (nh t l m sa thp). Nu tin hnh nhn thun ci tin n th s rt lu. Khi c th dng ging ny lm ging b ci tin v chn mt ging khc c u im m ging trn khng c lm ging i ci tin. em lai hai ging ny vi nhau th s c cc con lai vn c hng sn xut v cc c im c bn nh ging b ci tin nhng c thm c im ca ging i ci tin. Lai ci to Lai ci to l phng php lai mt ging c ci to (ging b ci to) vi mt ging khc (ging i ci to) bng cch lai lin tip. Dng con c ca ging i ci to phi vi con ci ca ging b ci to. Sau tip tc dng con c ca ging i ci to phi vi con ci lai (con ci ca ging b ci to ch dng mt ln). Khi t yu cu th c nh. Phn ln cc qun hp ging chp nhn lai bn th h lin tip vi mt c ging thun chng. Kt qu l to c cc con lai c h gene ca ging i ci to l chnh v ch c mt phn nh h gene ca ging b ci to. Th h con lai F4 c th c chp nhn l mt ging thun. Qu trnh ny cn nhiu nm v n phi ch i s sinh trng v pht trin ca cc con ci mi th h. Hin nhin, thnh cng ln nht s t c nu s dng cc con c ging tt nht c v ni ging v sc sn xut. Phng php ny thng c s dng rng ri khp ni trn th gii, trong trng hp khi mt ging tuy c mt s c im tt (c bit l tnh thch ng) nhng vn cn nhiu nhc im cn ci to. Nu tin hnh chn lc, nhn thun ci to th rt lu. Khi , c th dng ging ny lm ging b ci to (thng l ging cao sn nhp ni). Kt qu l hng sn xut ca ging b ci to s c thay i. Lai ci to v lai ci tin rt ging nhau v c th ni u l lai cp tin. Chng ch khc nhau ch, trong lai ci tin th cp tin h gene ca ging b ci tin cn trong lai ci to th cp tin h gene ca ging i ci to cho cc i sau. Lai cho (crossing for the production of new breed)
Lai cho l phng php dng hai hoc nhiu hn hai ging tin hnh lai. Sau chn lc v c nh cc i lai tt to thnh ging mi. Lai cho khng c mt cng thc c nh. 6.2. Cng ngh mi trong vic chuyn phi v thao tc phi 6.2.1. Chuyn phi (embryo transfer) K thut chuyn phi rt hiu qu trong vic di truyn cc gene qu. K thut ny c s dng qua mt s nm v cho php cc nh chn ging gia sc phn phi cht di truyn ca gia sc c gi tr nht ca h mc gia nh cng nh quc t m khng lm suy gim c s di truyn ca gia sc. S vn chuyn phi s thun li hn so vi vn chuyn cc ng vt sng. Cc nghin cu cho n nay cho thy rng phng php vn chuyn vt cht di truyn ny c th hn ch n ti thiu s xm nhp cc bnh truyn nhim vo cc qun th gia sc khc.
Thu nhn phi

Phng php phu thut v khng phu thut l hai phng php c s dng thu nhn phi cc i gia sc. Thu nhn phi bng phng php phu thut l phng php thu nhn phi sau khi m con vt. Cng c th git cht con vt, ct ly b phn sinh dc bn trong mang v phng th nghim di ra ly phi. Phng php khng phu thut c pht trin cho b v nga ci v cho kt qu nh phng php phu thut. Hin nay y l phng php ph bin c s dng thu nhn phi gia sc. Chng bao gm vic s dng mt ng thng Foley c 18-24 (gm hai ng lng vo nhau), cho php dn dung dch vo t cung v sau cho php dung dch t t cung quay tr ra vo mt vt cha chn lc. Mt qu bng nh gn cui ng thng c th c thi phng pha bn trong sng t cung ngn cn khng cho dch trn ra ngoi c t cung. Dung dch di ra thu c lng khong 20-30 pht, gn b phn bn trn. Phi c tch ra, a vo a petri v c nh gi phng i 75. Phi sng c phn loi v sp xp da vo s biu hin hnh thi ca chng. Phi sau khi nh gi phn loi c th em chuyn lun cho vt nhn ng pha (synchronized recipients) hoc em ng lnh s dng sau. Tt c cc phi sng c cho vo mi trng gi kh trng (DPBS b sung vi 0,4% BSA) theo s ch dn ca Hip hi Chuyn phi Quc t.
Chuyn phi
Phng php chuyn phi c s dng tng kh nng sinh sn ca ng vt ci. Hin nay y l phng php ph bin c s dng trong thc tin chn ging gia sc nhiu nc trn th gii. Tuy nhin
ng dng ca n trong vic nhn ging tru ch mi c bt u cch y hai thp k vi s ra i ca mt ch ngh con chuyn phi u tin. Cc phng php chuyn phi khc nhau c pht trin. C hai phng php chuyn phi: phng php phu thut v phng php khng phu thut. Phng php chuyn phi khng phu thut c mt s u im: t tn km, c th t t l th thai cao nh phng php phu thut. V vy phng php ny c s dng rng ri. Mc d nhiu cng c c thit k mt cch c bit cho vic chuyn phi bng phng php khng phu thut tru B c m t trong cc ti liu trong sut thp nin 1960 v u thp nin 1970 nhng mi cho n khi s p dng sng chuyn phi Cassou Al thnh cng, cui cng chng minh l c kt qu i vi vn c bit ny. Vo gia thp nin 1970, mt s sng chuyn phi cho gia sc cht lng by bn ch. Coultard (1991) (Vng quc Anh) m t phng php chuyn phi ca mnh bng cch s dng mt sng chuyn phi sa i. Sng ny c mt mng bc vi u kim loi v hai l thot hai bn. Khi thc hin chuyn phi, con nhn c gy t mng cng v sng lp t c a vo mng bc bng plastic rng hn pha ngoi. Chng c a vo n c t cung v sng c y xuyn qua mng bc pha ngoi. Vo nm 1983, Drost v cng s i hc Florida, Gainsville, M, thnh cng trong vic chuyn phi Tru. Nhm nghin cu ny chuyn mt phi tru 7 ngy tui vo con nhn v kt qu l ln u tin mt ngh con ra i bng phng php khng phu thut. S kin ny lm khuy ng s quan tm ng k ca mt s nc nui tru. Sau phng php ny c s dng n , Bungarie, Thi Lan, Pakistan, Malaysia (Misra, 1993), (Schallenberger, 1990), Vit Nam (Uoc, 1992), Philippines (Cruz, 1991), Nht Bn (Ocampo, 1988), Rp (Ismail, 1993) v Trung Quc (Wang, 1994). Drost cng l ngi i u trong vic nghin cu chuyn phi gia cc loi. Di s qun l ca Drost, nhm nghin cu chuyn mt phi tru sang mt b ci nhn. Tuy nhin, thai b sy giai on 2-3 thng tui (Drost, 1983). Chuyn phi bng phng php khng phu thut ln u tin Bungarie c thc hin vo cui nm 1984. Kt qu l th thai nhng con ci nhn sy thai vo thng th 3. Vo thng 4 nm 1986, ti vin Nghin cu Tru Shumen, Bungarie, mt th nghim nghin cu chuyn phi vi qui m ln gia Bungarie v M c tin hnh. Kt qu ca th nghim ny l ln th hai trn th gii v ln th nht chu u, 4 con ngh con chuyn phi ra i vo u nm 1987. Vo thng 2
nm 1988, nhm nghin cu Bungarie to c 3 ngh con chuyn phi (Alexien, 1988). Kt qu cc th nghim nghin cu u tin cho thy k thut chuyn phi p dng vi b khng cho cc kt qu ging nh tru do s sinh sn khc bit gia cc loi ny. Theo mt s th nghim, cc nh khoa hc Bungarie v M nh gi rng phi tru pht trin vi cng ln hn phi b trong 4 ngy u tin. V vy thi gian ti u cho s phc hi ca chng l gia ngy th 5 v ngy th 6 k t khi bt u ng dc thc s ca con ci cho. Chuyn phi vo thi gian ny em li s thnh cng ln hn. Chuyn phi khng phu thut c thc hin bng cch s dng mt sng chuyn phi thu nh xuyn qua c vo sng t cung. Cc con nhn cng lc c kim tra s c mt ca mt th vng hot ng (CLcorpus luteum). Mng cng c gy t l gim n mc ti thiu sc cng. Phi chuyn c ht vo mt cng rm 0,25 ml trong mt ct trung tm cha 20 ml mi trng gi (holding medium) nm gia hai ti kh. Cng rm c lp vo sng chuyn phi v mt mng bc vi u kim loi c lp qua nh. Sau mt mng bc v sinh c qun trn nh trnh bt k s nhim trng no t h vi khun m o. By gi sng chuyn phi c a xuyn qua m o n ngoi ming t cung. Ri th mng bc v sinh c xuyn qua v sng chuyn phi c a t t qua c v thn t cung n trn 1/3 sng t cung, cng pha vi bung trng mang th vng. Piston ca sng c y chm chm t phi vo sng t cung v sng c rt ra t t. Trong chuyn phi bng phng php khng phu thut, bc quan trng nht l a mt dng c vo nh sng chuyn phi n c v sng t cung. S s dng khng cn thn cc dng c nh th s lm tn thng c cng nh mng t cung v gy chy mu. Do ch di chuyn cc dng trn khi bit v tr ca chng v qu trnh a cc dng c vo t cung lun c kim tra v iu chnh qua trc trng. nc ta, ti Vin Cng ngh Sinh hc thuc Vin Khoa hc v Cng ngh Quc gia, vo ngy 13 thng 4 nm 1997, mt con d c Vit Nam sau 4 thng 23 ngy mang thai t phi d Php c gi ng lnh sinh ra con d ci Php ging Alpin mu trng tuyn thun chng. 6.2.2. Cc thao tc phi Chia tch phi (embryo spliting) Vi k thut ny c th cho ra nhiu phi t mt phi ban u, to ra hng lot cc c th ging ht nhau v mt di truyn hay ni cch khc l to nn mt dng v tnh.
C hai phng php chia tch phi: phng php dng kim v phng php dng dao ct. Phng php dng dao th n gin hn v d dng hn nhiu so vi phng php dng kim. thc hin chia tch phi cn phi c dung dch nui phi, knh hin vi soi ngc Olympus, thit b vi thao tc iu khin dao ct, kim gi c nh phi... Trc ht cho phi dng chia tch vo a petrie cha dung dch nui cy; c nh phi bng kim gi; iu chnh thit b vi thao tc iu khin dao ct theo hng thng ng t trn xung hay theo hng nm ngang sao cho li dao t ng vo gia khi t bo phi; ct phi thnh hai, ch thao tc nhanh, dt khot v chnh xc; chuyn phi sau khi tch vo dung dch nui mi nui cy khong 2-3 gi trc khi chuyn vo vt nhn c gy ng dc ng pha; cng c th nui cy v tip tc chia tch lp li thu nhn c nhiu phi hn. Phi c s dng chia tch t nht l giai on 5-7 ngy sau khi th tinh (i vi b). Nu s dng phi giai on cui phi du hay u phi nang khi khi t bo ln v cc t bo cha bit ha th kt qu chia tch phi ni chung l s tt hn. Nu chia tch phi giai on phi du th ch vic chia khi mm phi thnh hai phn u nhau. Cn nu chia tch phi giai on phi nang th ngoi vic tch ni phi b th cn phi tch phn ngoi phi b. Nu tch phi giai on mun hn, lc cc t bo bit ha th t l to nn nhng c th ton vn l thp.
Sinh thit phi

Sinh thit t phi sinh i cng trng c th xc nh c gii tnh v cc c tnh di truyn ca dng v tnh. C th ht ra mt t t bo t phi xt nghim hoc dng dao ct mt phn ca phi. Chuyn ghp nhn (nuclear transplantation) Phng php chuyn ghp nhn to nn cc dng v tnh thnh cng nhiu loi gia sc nh cu, b, nga, ln, d. y l mt phng php hin i nhm chuyn nhn ca mt t bo phi sm vo mt t bo trng tch nhn i to nn t bo lng bi (hp t) v pht trin thnh phi. Trc ht, gy siu bi non, th tinh, ri thu nhn phi tt nht l giai on phi du; tch khi t bo phi du thnh tng t bo (2n) ring l; cy mi t bo (2n) ny vo mt t bo trng cha th tinh c loi b nhn t trc; dung hp t bo (2n) vi t bo trng cha th tinh khng nhn bng xung in; cc t bo trng ghp nhn c nui cy in vitro hoc a vo nui vt nhn trung gian thng l th hoc cu; sau chuyn cc phi pht trin ny vo cc vt nhn c gy ng dc ng pha. Phng php ny em li hiu qu cao trong nhn ging, trong ci tin di truyn cc ging vt nui.
6.3. Cng ngh sinh hc trong vic chuyn gene to ging vt nui Mc ch ca cng tc chn ging v nhn ging l ci tin tim nng di truyn ca vt nui...nhm nng cao nng sut, hiu qu sn xut nng nghip. Trong cng tc nhn ging c truyn ch yu s dng phng php lai to v chn lc ci to ngun gene ca sinh vt. Tuy nhin, do qu trnh lai to t nhin, con lai thu c qua lai to v chn lc vn cn mang lun c cc gene khng mong mun do t hp hai b nhim sc th n bi ca giao t c v giao t ci. Mt hn ch na l vic lai to t nhin ch thc hin c gia cc c th trong loi. Lai xa, lai khc loi gp nhiu kh khn, con lai thng bt th do sai khc nhau v b nhim sc th c v s lng ln hnh thi gi b v m, do cu to c quan sinh dc, tp tnh sinh hc... gia cc loi khng ph hp vi nhau. Gn y, nh nhng thnh tu trong lnh vc DNA ti t hp, cng ngh to ng vt chuyn gene ra i cho php khc phc nhng tr ngi ni trn. N cho php ch a nhng gene mong mun vo ng vt to ra nhng ging vt nui mi k c vic a gene t ging ny sang ging khc, a gene ca loi ny vo loi khc. Cng ngh to ng vt chuyn gene l mt qu trnh phc tp v nhng loi khc nhau c th khc nhau t nhiu nhng ni dung c bn gm cc bc chnh sau: 6.3.1. Tch chit, phn lp gene mong mun v to t hp gene biu hin trong t bo ng vt Mt gene ngoi lai trc khi c chuyn vo genome ca t bo vt ch to ra ng vt chuyn gene phi c phn lp v tinh ch hay ni cch khc l n phi c to dng. Cc cng c s dng to dng bao gm cc enzyme c bit c hot tnh ct v ni DNA (enzyme hn ch v ligase), cc mu d (probe), vector v t bo vt ch. T bo vt ch thng c s dng l t bo vi khun E.coli v vector thng c s dng l plasmid. Vic tch chit mt gene ring l l rt phc tp bi v DNA mu cha hng triu gene. Do thc hin iu ny, DNA mu cha gene mong mun v vector plasmid phi c ct bi cng mt loi enzyme hn ch. Cc on DNA mu sau khi c ct c mang gene mong mun s c xen vo vector plasmid v cc u ca cc on DNA mu ny v cc u ca vector plasmid s c ni vi nhau nh ligase to thnh plasmid ti t hp. Sau cc plasmid ti t hp c bin np vo cc t bo vi khun E.coli v cc t bo vi khun tin hnh sinh trng. Vo thi im ny, cc t bo vi khun cha plasmid mang gene mong mun s c pht hin bng mu d. Chng c nui cy trong mi trng thch hp sinh trng pht trin to ra hng triu bn sao ca vector cha
gene ny. Vector cha gene ny s c tch ra khi t bo vi khun v gene mong mun s c tch chit. Phng php ny c th to ra hng triu bn sao ca gene mong mun m khng b nhim bi cc gene khc. Gene chuyn c ngun gc t genome ny cha cc on exon m ha v cc on intron khng m ha . Ngi ta cng c th phn lp c gene mong mun t sn phm biu hin ca n nh mARN hoc protein. T mARN di tc ng ca enzyme sao chp ngc s tng hp ra DNA b sung mch n (ss cDNA), tip theo l DNA b sung mch kp (ds cDNA). DNA b sung (cDNA) ny khc vi DNA gc l khng cha cc intron m ch bao gm cc exon. Hoc t sn phm protein, c th suy ra trnh t nucleotide ca gene cu trc trn c s trnh t cc acid amin trong phn t protein. Sau c th thit k cp mi (primer) d tm on gene mong mun. Gene cu trc mun hot ng biu hin ra protein m n qui nh trong h thng t bo nht nh th phi c promoter thch hp vi h thng m n hot ng. Promoter t bo ng vt c ngun gc hoc t ng vt nh methallothionein (MT), thymidine kinase, -actin, amylase, insulin, -lactoglobulin, adiposite P2 ... hoc t virus ng vt nh Simian virus (SV40), Rous sarcoma virus... 6.3.2. To c s vt liu bin np gene ng vt c v th giai on bin np gene thch hp nht l trng giai on tin nhn (pronucleus), giai on m nhn ca tinh trng v trng cha dung hp (fusion) vi nhau. giai on ny t hp gene l c c hi xm nhp vo genome ca ng vt nh s ti t hp DNA ca tinh trng v ca trng. Do t bo phi cha phn chia v phn ha nn t hp gene l c bin np vo giai on ny s c mt tt c cc t bo k c t bo sinh sn ca ng vt trng thnh sau ny. i vi ng vt c v, trng chn c thu nhn bng phng php s dng kch dc t theo chng trnh c xy dng cho mi loi hoc bng phng php nui cy trng trong ng nghim. Sau th tinh nhn to to ra trng tin nhn. 6.3.3. Chuyn gene vo ng vt Vic t c mc ch ca vic to ra ng vt chuyn gene i hi s pht trin ca cc phng php c hiu qu chuyn gene mong mun vo phi. Hin nay c nhiu phng php chuyn gene khc nhau ang c s dng to ng vt chuyn gene: vi tim, chuyn gene bng cch s dng cc t bo gc phi, chuyn gene bng cch s dng vector virus... 6.3.4. Nui cy phi trong ng nghim
T bo trng tin nhn sau khi vi tim c nui cy trong ng nghim pht trin n giai on phi du hoc ti phi. giai on ny mng trong (pellucida) b bong ra v phi c th lm t c d con. Nhng phi ny c cy chuyn vo con nhn c gy cha gi (pseudopregnant) pht trin thnh c th con. 6.3.5. Kim tra ng vt c sinh ra t phi chuyn gene khng nh ng vt c c chuyn gene l vo hay khng, ngi ta phi kim tra xem gene l c xm nhp c vo b my di truyn ca ng vt trng thnh hay khng v sn phm ca gene l c c tng hp ra hay khng. i vi vn th nht ngi ta s dng phng php lai phn t trn pha rn (Southern blot, Northern blot...) hoc PCR. i vi vn th hai, pht hin protein do gene l tng hp ra ngi ta s dng phng php Western blot, ELISA hoc k thut min dch phng x (RIA) pht hin protein l trong ng vt. Theo di cc th h sau ca ng vt chuyn gene (F 1, F2, F3, ...) xc nh gene l c di truyn hay khng. 6.4. Gia sc chuyn gene, nhn bn v tnh ng vt v mt vi thnh tu mi 6.4.1. Gia sc chuyn gene
Gia sc chuyn gene hormone sinh trng

Trong hng ny ngi ta tp trung ch yu vo vic a t hp bao gm gene cu trc ca hormone sinh trng v promotor methallothionein vo ng vt. Cho n nay ngi ta a thnh cng gene ny vo th, ln v cu. Kt qu l nhng ng vt chuyn gene ny khng to ln nh chut. Tuy nhin c, trong trng hp ln chuyn gene hormone sinh trng lng m gim i ng k (gim t 28,55mm xung 0,7mm) v hiu qu s dng thc n cao hn. Australia, ln chuyn gene hormon sinh trng c tc ln nhanh hn i chng l 17%, hiu sut s dng thc n cao hn 30%. Cc nh khoa hc Granada (Houston, Texas) to ra c b chuyn gene tip nhn estrogene ngi (human estrogene receptor) c tc ln nhanh. Cc nh khoa hc y thnh cng trong vic a gene hormone sinh trng ging insulin b (bovine insulin like growth hormone) vo gia sc to ra ging gia sc tht khng dnh m.
Gia sc chuyn gene cung cp cc dc phm cha bnh

Nh chng ta bit, nhiu protein dc phm qu khng th sn xut qua con ng vi sinh hoc sinh vt bc thp, do nhng sinh vt ny khng c h enzyme to ra nhng protein c cu to phc tp. nh s dng tuyn sa ca ng vt bc cao sn xut ra protein qu ln u tin c Clark (1987) xut. Ni dung ca k thut ny l gn gene cu trc vi -lactoglobulin (l promoter iu khin s biu hin ca gene tuyn sa). Khi a t hp c cha promoter -lactoglobulin vo cu, chut, Clark thy chng biu hin rt cao tuyn sa (Hnh II.3. ). Cho n nay rt nhiu protein dc phm qu v ang c nghin cu sn xut qua tuyn sa ca ng vt nh: - a1- antitripsin v yu t lm ng mu IX (Blood Clotting Factor IX) ca ngi c tit ra trong sa cu vi nng l 25mg/ml. - Cht hot ha plasminogene m ngi (Human Tissue Plasminogene Activator) lm tng ng mu c tit ra sa d. - Gene urokinase ngi c a thnh cng vo ln v tit ra tuyn sa nh gene khi ng alpha-casein b. - Protein C ngi c to ra t sa ln chuyn gene...
Gia sc chuyn gene khng bnh

n nay ngi ta bit c mt s gene c kh nng khng bnh ca vt nui. Tim gene Mx vo ln to ra c ging ln min dch vi bnh cm. Ngi ta cng thnh cng trong vic tim gene IgA vo ln, cu, m ra kh nng to c cc ging vt nui min dch c vi nhiu bnh... Gia sc chuyn gene cho nng sut cao v cht lng sn phm tt Trong hng ny ni bt l nhng nghin cu nng cao cht lng sa b, sa cu bng cch chuyn gene lactose vo cc i tng quan tm. S biu hin ca gene ny c iu khin bi promoter ca tuyn sa. Trong sa ca nhng ng vt chuyn gene ny, ng lactose b thy phn thnh ng galactose v ng glucose. Do vy nhng ngi khng quen ung sa cng c th s dng c sa ny m khng cn qu trnh ln men. Hin ti ngi ta ch ti vic a mt s gene ca vi sinh vt vo c th ng vt. Tin b ni bt nht trong hng ny l a gene m ha enzyme chu trch nhim tng hp axt amin cystein vo cu. Cystein l acid amin c tng hp t serine nh hai enzyme l: serine transacetylase va O-acetylserine sulfahydrylase. Hai gene chu trch nhim tng hp hai enzyme ny l Cys E v Cys K. Cystein l acid amin c bn rt quan trng trong s pht trin ca lng. Nhng c gng b
sung acid amin ny vo thc n u khng t kt qu do chng b phn hy trong ng tiu ha ca ng vt. Bi vy, nu a c gene tng hp cystein vo c th ng vt s lm tng nng sut lng ln rt nhiu. Cc nh khoa hc Australia dng gene tng hp cystein c ngun gc t vi sinh vt (E.coli v S. typhimurium) a vo cu. Hai gene Cys E v Cys K phn lp t hai chng vi sinh c gn vi nhau thnh mt on DNA, sau c gn tip vi promoter methallothionein. chut c chuyn t hp gene ny th hu ht cc c quan u c mt t
Hnh II.4. S qui trnh sn xut protein thng qua tuyn sa
hp v lng lng tng ln rt nhiu. S biu hin chut v cu ging nhau nn trong tng lai khng xa s to ra c ging cu chuyn gene cystein vi nng sut lng tng ln rt nhiu ln. Gia sc chuyn gene cung cp cc ph tng ghp thch hp phc v y hc Yu t cn bn trong vic to ra vt nui chuyn gene cung cp ph tng thch hp cy ghp cho cc bnh nhn l ngn cn s loi thi th ghp nh s hot ha cc nhn t b sung thuc h min dch ca ngi. Cc nh khoa hc to ra ln chuyn gene biu hin gene m ha cc nhn t ngn cn s b sung ca ngi (human complement inhibitory factors) nh nhn t lm tng nhanh s phn hy (decay-accelerating factor) (Rosengard, 1995). Mc tiu ny ang c tip tc nghin cu. 6.4.2. Nhn bn v tnh ng vt v mt vi thnh tu mi Vo nm 1996, Wilmut, Campbell v cng s thnh cng trong vic nhn bn to ra ch cu Dolly. Cc nh nghin cu s dng cc loi t bo cho nhn l t bo a phi (9 ngy sau th tinh), t bo thai (26 ngy sau th tinh) v t bo tuyn v ca mt cu ci 6 nm tui ang giai on 3 thng cui ca thai k. Gii bit ha bng cch a nhn v trng thi tnh, pha G0, G1 hay M: gim t l huyt thanh trong mi trng nui cy (t 10% xung cn 0,5%). Kh nhn non bo, hot ha non bo v dung hp non bo vi t bo tuyn v bng xung in. V kt qu thu c l mt phi. T 227 phi thu c bng phng php ny c a vo nui cy vi trng c tht li ca cc cu ci mt en. Cc phi thu c giai on phi du hay phi nang li c chuyn vo t cung ca cc cu ci nhn khc. Sau 5 thng, ch c 1 phi pht trin thnh cu con Dolly bnh thng. (Phn ny xin xem thm Chng XI: Cng ngh sinh hc trong vic bo tn v pht trin ngun gene qu him. V ng vt, trang 205) 7. Vector virus sng trong to vaccine th y ti t hp 7.1. Virus u ma ti t hp Virus u ma l thnh vin ca h poxvirus. DNA genome ca virus ny dng si kp c kch thc khong 187 kb v m ha 200 protein khc nhau. DNA virus u ma ti bn trong t bo cht ca t bo b nhim. S ti bn c th xy trong t bo cht hn l trong nhn bi v DNA virus u ma m ha cc gene DNA polymerase, ARN polymerase v cc enzyme xc tc cho qu trnh to m cap, methyl ha, gn ui polyA vo mARN. Virus ny c th xm nhim vo cc ng vt c xng sng v ng vt khng c xng sng. Virus u ma c dy vt ch ln, biu th c im tt mc phn t v thng l virus n ha. V vy n l mt vector vaccine. Cc c
tnh quan trong ca mt vector vaccine l pht tn v biu hin cc gene c to dng m ha cc khng nguyn m n n cc khng th trung ho chng li cc tc nhn gy bnh. Tuy nhin genome virus u ma l rt ln v thiu cc v tr ct hn ch, v vy n khng th xen thm vo cc on DNA mt cch trc tip. iu cn thit l cc gene i vi cc tc nhn c hiu phi c a vo genome virus bng ti t hp tng ng (homologous recombination) in vitro. Mt s gene khng nguyn c xen thnh cng vo genome virus u ma v sau biu hin cc t bo ng vt nui cy. Cc khng nguyn ny bao gm protein G ca virus di, khng nguyn b mt ca virus vim gan B, protein b mt ca virus Sindbis, protein HA v NP ca virus cm, protein N v G ca virus vim ming v glycoprotein ca virus herpexs simplex. S s dng cc vaccine vector c to ra t virus u ma cho php cc c th chng nga c kh nng chng li mt s bnh khc. Kt qu ny c c l do s dng mt virus u ma ti t hp mang cc gene to dng m ha mt s khng nguyn khc nhau. So vi cc loi vaccine khc, vaccine virus ti t hp sng c mt s c im thun li sau y: - Virus ny c th biu hin cc khng nguyn ng tin cy mt dng gn ging vi s xm nhim t nhin. - Virus u ma c th ti bn trong vt ch, do khuych i lng khng nguyn hot ha s gii phng khng th t cc t bo B v kch thch s sn sinh cc t bo T. - S xen cc gene khng nguyn vo mt hoc nhiu v tr trong genome virus u ma lm gim nhiu hn tnh c ca n. Trong khi nhiu cng trnh nghin cu pht trin vaccine vector virus sng c tin hnh vi virus u ma, cc virus khc nh adenovirus, varicella-zoster virus cng ang c nghin cu nh l cc th truyn vaccine c tim lc. 7.2. Adenovirus Adenovirus gy ra nhiu bnh khc nhau cho ngi v ng vt. Chng ang c nghin cu lm vector biu hin cc protein khng nguyn sn xut cc vacccine virus ti t hp chng li cc bnh tt con ngi v ng vt. DNA geneome ca adenovirus c kch thc 36 kb, cha hai vng xen DNA ngoi lai vo: vng E3 v vng E1. Vng E3 khng c kh nng can thip virus nhn ln trong t bo nui cy v a s l hay b mt. DNA ngoi lai c xen vo mt plasmid vi khun v sau c ci vo genome adenovirus. Adenovirus ny c ly nhim vo t bo ng vt c v. Adenovirus c nhng thun li l d dng nhn
ln trong t bo b ly nhim v genome ca n rt bn. Mt khc mc biu hin ca gene khng nguyn ngoi lai rt cao. Hn na cc protein khng nguyn cn c th ci bin sau qu trnh phin m. 8. Cng ngh sinh hc vi vic nui trng thy sn Nhu cu v lng thc, thc phm tng ln cng vi tng dn s, nn i protein tr nn trm trng. Mt nc, c bit l bin c xem l mt trong nhng ni c kh nng gii quyt kh khn , trong thy sn gp phn cung cp ngun protein, lipid, glucid,cho con ngi. Do khai thc thy sn qu mc, hot ng pht trin kinh t, th ha, pht trin cng nghip, ph rng, gy nhim v ph v cc h sinh thi: rn san h, thm rong to, rng ngp mn, nhim ngun nclm mt ni c tr v bi ca nhiu loi thy sn, sn lng t nhin c chiu hng gim st. Theo FAO sn lng khai thc thy sn trn th gii trong tng lai ch gi mc hin nay (85-90 triu tn/nm). Trc tnh hnh , vic nui trng thy sn l mt gii php nhm b sung v tng cng sn lng thy sn trong thi gian ti. nc ta, vic nui trng thy sn v ang tr thnh mt hng trng im. Vic ng dng cc thnh tu CNSH l mt bin php nhm to ging mi, nng cao nng sut v cht lng sn phm. 8.1. Mt nc nui trng thy sn Mt nc trn th gii chim mt din tch kh ln: 360 triu km2 trong s 510 triu km2 ca ton lc a. M u c nc th c cc sinh vt sinh sng. T cc ao h, m ph, sng sui, mt bin, nc ngt, nc l hay nc mn, qua qu trnh pht trin c th sinh vt thch nghi vi mi trng v sinh trng pht trin mt cch bnh thng trong iu kin sng . Do , thc vt nc dng carbon ng ha trong kh quyn l 15,3.1010 tn, trong lc ton b thc vt hp th carbon cng ch 17,4.1010 tn. Nhn dn th gii cng bit s dng mt nc khng l nui trng thy sn. Nm 1995, Vit Nam c xp th 13 v s dng mt nc nui trng thy sn. Hin nay c xp th 5 trong khu vc Thi Bnh Dng v din tch nui trng trong nm 2000 ln xp x 135.000 ha. Din tch nc ngt tp trung nhiu nht Nam B (55,07%), Bc B chim 24,15%, Trung B l 20,78%. Thnh phn cc loi c nui cng rt phong ph, ring c ni a c 28 loi, c nhp ni l 11 loi vi 7 dng c khc nhau. (thng tin t b Thy sn nm 1996). Nhiu i tng khc nh gip xc (tm, cua,) hai mnh v, c, ba ba, i mi, rong tocng c a vo nui nhng qui m khc nhau. 8.2. Cng ngh sinh hc trong sn xut ging thy sn
Sn xut ging nhn to tr thnh nhu cu bc thit trong nui trng thy sn. pht trin m rng nui cc i tng c hiu qu, vn ging tr thnh mt mt xch quan trng. Cc thnh tu v sinh sn nhn to trn cc i tng thy sn cc nc khc nhau trn th gii cng nh Vit Nam l rt to ln. T khi cc nh khoa hc nc Nga cho sinh sn nhn to thnh cng trn c m ra trin vng to ngun ging nhn to phc v vo vic nui trng ngy cng m rng khp ni trn th gii. Hawaii, M t nm 1976, cc nh nui trng cho sinh sn nhn to trn mt s i tng c bin. mt s nc nh Nht Bn, Nga, M, Canada ng dng tnh di c hai chiu, cho sinh sn nhn to trn c hi, ging c th li t nhin, sau c ln quay vo t lin v t chc nh bt. Hnh thc ny cng c p dng i vi c chnh l i tng c qu him, lc trng thnh sng nc ngt, khi di c ra bin. Nga v Iran cng cho sinh sn trn c tm, a vo th t nhin v khai thc khi chng ln ln. Trong khi Na Uy l nc sn xut ging c hi phc v nui qui m cng nghip bng nhng lng bin, nm 1995 sn lng t 220.000 tn, gi tr 1 t USD. Trung Quc l nc c nhiu loi c nc ngt nh: c trm en, trm c, m hoa, c chp, cho sinh sn nhn to thu c mt s lng ln c ging, phc v ngh nui rt pht trin. Nht Bn, cc i tng tm he, trai ngcc sn xut ging phc v cho vic nui. khu vc ng Nam , tp trung nghin cu nhiu i tng c nc ngt, cho sinh sn nhn to em li nhng kt qu kh quan. Ring Thi Lan, sn lng c nc ngt t nui trng nm 1993 l 161.600 tn, nui 3 loi chnh l c r, c tr, c m vinh. Philippines, cho sinh sn nhn to thnh cng nhiu i tng nui nh c trm, c m, c bit l c mng bin c sinh sn nhn to a vo nui v t sn lng ln. Vit Nam vic cho sinh sn nhn to thnh cng trn c m hoa t nm 1963, m ra mt trin vng trong vic sinh sn nhn to trn c nc ngt. Tip theo nhng nm 1967-1970, v c bn nc ta sinh sn thnh cng hu ht cc ging c nui, ch yu bao gm c a phng v c nhp ni. Nm 1996, c nc ta c 375 tri ging, hng nm sn xut 4-5 t c bt bng phng php s dng hormone HCG, hypophyce v ng c 800-900 triu c ging. T nm 1990-1998, Nam B cho thnh cng c tra, c ba sa, c bng tng, c tr lai, c r phi, tm cng xanh. Ngoi ra chng ta cn cho sinh sn nhn to thnh cng nhiu i tng nc ngt c gi tr khc nh ba ba, ch. Hng nm c nc ta sn xut hng trm triu con ging thuc i tng thy sn nc ngt a vo phc v cho nhn dn.
Vic tip tc a dng ha cc i tng nui a vo nui, tn dng cc mt nc, chng ta cho sinh sn nhn to thnh cng nhiu i tng nc mn. Vic cho thnh cng i tng tm s ( Penaeus monodon) v a vo nui qui m c nc, tip theo thnh cng phng php ct mt v cy tinh lm cho hiu qu c nng cao. Sau nhiu i tng nui nc l, nc mn khc cng cho thnh cng nh c chm, c bp, c hng, ip qut, trai ngc, c nga, bo ng, hi sm, v.v. Vic hiu bit v s phn b, c im sinh hc, sinh thi ca chng cho php chng ta tin hnh sinh sn thnh cng v ng nui giai on u trng thnh ging vi t l sng cao v c cht lng tt m rng thnh qui m c nc. V d: tm s: c nc c trn 3000 tri cho t bc n nam, ring Khnh Ha c n 1000 tri. Qui trnh cho trn tng i tng cng khc nhau. Chng hn, i vi c, ngi ta theo di trn cc giai on pht trin ca bung trng ca c ci v tinh so ca c c, trong giai on ny c th tim hormone sinh dc kch thch. Sau khi cho th tinh xong, ngi ta cho vo trong b p, lc ny bt u theo di qu trnh phn chia v t l n ca trng, qu trnh ny ph thuc rt nhiu vo k thut cng nh kinh nghim. Trng n v tri qua mt s giai on. C bt khi n giai on dinh dng, ngi ta cho chng n cc loi thc n ph hp vi la tui ca c bt c t l sng cao. Cc yu t mi trng nh nhit , mn, pH,cng c vai tr quan trng khng km. i vi gip xc, con ci bt trong t nhin m trng. Cho con c v con ci vo chung mt b c kch thc thch hp, nu con ci trng chn mui, ngi ta kch thch cho chng . Sau khi th tinh, trng a sang b p, y trng phn chia. Tri qua mt s giai on bin thi v n giai on ging mt khong 15 ngy, ty thuc vo tng giai on, ngi ta cho n to, Artemia Sau y l m hnh ca Thi Lan. tm m c mua t cc ngun cung cp ging, sau , cho vo b t 50-60 con ci v 20-50 con c trong b xi mng ng knh rng 4 m (cho n bng polychaete-Nercis spp v mc ng), con ci mang trng a sang nhng b 500 lt bng si thy tinh, mi con ci mt b. Nhng con ci khng c trng c kch cho bng phng php ct mt.Mt con khong 800.000-1.200.000 u trng. Nc trong cc b c kim tra cht lng hng ngy, cc yu t kim tra l pH, mn, nhit , ammonium (NH4+). Cc b u c sc kh. Sau khi th tinh, u trng tch khi v trng v lng xung. Dng ng ht u trng vo b ng c th tch 4m3, giai on u zoea mysis cho n to Chaetoceros trong 4 ngy, giai on PI.4-PI.5 (post larvae) cho n Artemia. Tm nui t 15-20 ngy, xut b bn.
B lc nc
B ng
B x l nc thi
M hnh ng Tm s Ngoi vic cho thnh cng cc i tng nui kinh t, cc nghin cu nhng php lai kinh t ng dng tnh u th lai th h F1 cng c p dng. Nm 1979, Vin Nui trng Thy sn 1 cho lai kinh t gia c chp trng Vit Nam vi c chp lai nhp ni, c lai ngc ln lai xui u cho u th lai, t l sng ca th h F 1 t rt cao t 44-80% (trung bnh l 62%), trong khi c chp trng t l sng trung bnh l 50,35%. u th lai ln nhanh hn hn b m t 180-230%. Cc nm sau, cc i tng khc cng c p dng lai kinh t nh c r Phi loi Oreochromis mosambicus vi loi O. niloticus, c tr loi Clarias garienpirus vi loi C. fuscus v loi Clarias garienpirus vi loi C. macrocephlus, c m trng Vit Nam vi c m trng Hoa Nam (Trung Quc) Ngoi cc loi c nui, ngi ta cng ch trng n cc loi c cnh nui trong cc aquarium. Theo b Thy sn (1996), nc ta c n 83 loi c cnh c nui. 33 loi c khc c mu sc p cha c a vo nui v 35 loi nhp ni c ngun gc khc nhau t chu M, chu Phi, n , Malaysia v ng Nam . Tng s loi c nc ngt dng lm c cnh ln n 151 loi. Nhiu loi trong s cho sinh sn thnh cng. Nhiu a phng nh thnh ph H Ni, thnh ph H Ch Minh c nhiu gia nh nui v kinh doanh c cnh. Trong my nm gn y, nhiu loi c bin cng c a vo nui trong cc aquarium phc v khch du lch, nht l cc rn San h c mu p (c n trn 1000 loi). Nhiu loi trong bc u cho sinh sn nhn to, nh cc loi ca ging c khoang c l loi sng cng sinh vi hi qu. Vic bc u ng dng cc thnh tu cng ngh sinh hc hin i vo lnh vc sinh sn nhn to c mt s kt qu. Vic ng dng k thut di truyn bc u c th nghim trn c hi (Salmon) bng phng php chuyn gene sinh trng ngi cho ta thy c nui sau 4 thng c sc tng trng nhanh hn c i chng. Vit Nam Nguyn Vn Cng v cng s (1999) nghin cu phng php chuyn gene hormone sinh trng ngi trn c vng v c chch cng cho kt qu tt, m ra trin vng ng dng cho to ging thy sn trong tng lai.
V vic nghin cu cng ngh sinh hc trong nui trng cc loi rong to. Sau khi s dng cc dch huyn ph loi to lc n bo Chlorella, Emerson R., Lewis C.M., 1941; Dulton H.J., Manning W.M., 1941 pht hin thy c mt s vi to c th tng sinh khi ca mnh trong mi trng vi s lng 1-2 triu t bo/ml cht lng, sau vi gi mt huyn ph t ti 50-100 triu t bo/ml huyn ph, sinh khi ny cha ti 50% protein th. u thp k 40 ca th k trc nhiu thc nghim nui cy vi to qui m ln c trin khai c. Nm 1953, cc nh nghin cu c trm Kohlensffbiologische Froschungstation (vng Essen CHLB c) nghin cu kh nng dng CO2 ph phm ca nh my vng Rubirdder nui trng to Chlorella v mt loi to lc n bo khc l Scenedesmus acutus. u nm 1970, chnh ph c m rng vic h tr nui trng to nhiu nc ang pht trin nh n , Pru, Thi Lan, u thp k 60 ca th k XX, vic nui trng to Spirulina, mt loi to c nh N2 li cun s quan tm ca cc nh khoa hc vi cng trnh nghin cu tin phong ca Clement G. v cng s. Nm 1975, Oswald W. J. v cc cng s trng i hc California cng b cc kt qu nui trng to qui m ln va thu nhn sinh khi va x l nc thi. Sau ny c nhiu nc nh Mexico, Nht Bn, Thi Lan, Php, Slovaki, v c Vit Nam cng tin hnh nghin cu v nui trng to Spirulins maxima v S. platensis. Cc nghin cu v to lam c nh m nh Anabaena sianensis, Calothrix spp., Hapalosiphon spp., c tin hnh n , Myanmar, Trung Quc, Philippines, Thi Lan, Vit Nam v a vo ng dng trong sn xut. Cc nh khoa hc Israel phng th nghim Cng ngh Sinh hc To t ra mt chng trnh nghin cu nhm phn lp cc loi to lam sng t do hay cng sinh trong rung la, nghin cu iu kin sinh trng ti u ca chng. V to ln, nhiu nc trn th gii hin nay ang nui trng v sn xut qui m ln dng trong nhiu lnh vc cng nghip v thc phm nh rong m (Sargassum), cc loi agarophyte, rong sn (Kappaphycus alvarezii Doly). Cc nh khoa hc nghin cu su v c im sinh l sinh ha xc nh thi v nui trng v phng php ch bin thch hp vi cc vng sinh thi khc nhau trong thy vc. C nhng nghin cu su nhn ging bng bo t hoc to nhng loi mi nh tch protoplast dung hp, tch t bo t Gracilaria nhm tm ra nhng loi agarophyte va c nng sut cao va c phm cht tt.
8.3. Cng ngh sinh hc trong k thut nui trng thy sn nng cao sn lng trong nui trng thy sn, ty theo tng i tng c qui trnh v hnh thc nui thch hp. Nhn chung c 4 hnh thc nui c bn sau: - Nui qung canh - Nui qung canh ci tin - Nui bn thm canh - Nui thm canh Vic ng dng s hiu bit vo cc hnh thc nui trn l quan trng nhm em li hiu qu cao, chi ph thp, tn dng c iu kin t nhin v x hi ca tng quc gia v tng a phng. Ty thuc vo hon cnh ca a phng v trnh ca ngi nui m c cc m hnh nui khc nhau.V d: Khnh Ha hu ht ng dng hnh thc thm canh v m hnh nui tm khp kn, cn min Nam th dng nhiu hnh thc. Nui trng thy sn cn kt hp vi chn nui, lm nng nghip, lm nghip, hnh thnh cc m hnh VAC, VACR, v.v. Cc m hnh ny em li hiu qu kh tt. Ngoi ra c cc m hnh khc nh nui kt hp la-c, la-tm hoc tm-mui, Gn y hnh thc nui tm trn triu bng cch xy dng cc ao nui trn cc bi ngang, nui tm trn ct, cng em li hiu qu nh Ninh Thun, Qung Ngi, Tha Thin-Hu, Qung Tr, H Tnh, Vic tn dng cc h ao sng sui nc ngt, cc h treo nc u ngun th c hoc nui bng b em li hiu qu ln. cc thy vc nc l v mn cng c trin khai nui trng nh tm s, tm hm, vm xanh, c hng, ngc trai, rau cu tht, rau cu cc, rong sn. Cc i tng ny nui ring l c v nui kt hp c s dng trit mt nc v x l cht thi trong mi trng nui. 8.4. ng dng cng ngh sinh hc trong sn xut thc n nui trng thy sn Sn xut thc n l gii quyt khu quan trng trong nui trng thy sn qui m cng nghip, phc v trong sinh sn ging nhn to. Ngi ta tm ra ngun thc n t giai on bo xc (cyst) ca to, Artemia, Rotifer nhm ch ng c ngun thc n ch ng cho tng giai on pht trin u trng va tit kim c thc n li em li hiu qu cao. Ngoi ra, cc giai on u th naupli ca nhiu loi thc vt v ng vt ph du cng c sn xut lm thc n cho u th tm, c, Trong sn xut thc n nhn to, ngi ta sn xut Chaetoceros spp. lm thc n cho Tm s giai on mysis n post larvac 4 ngy tui (PL 4) bng mi trng KNO3 100 mg/m3, Na2HPO4 10 mg/m3, Na2SiO3 5 g/m3, FeCl2 2,5 g/m3. Hin nay nhiu c s nghin cu nc ta u c kh nng sn xut sinh khi cc loi to, Rotifer nh Vin Hi Dng hc
Nha Trang, i hc Thy sn Nha Trang, Trung tm Nghin cu Thy sn III dng lm thc n trong nghin cu sn xut ging. Hu ht cc tri sinh sn nhn to tm s u s dng to v Artemia lm thc n. to dng c r phi n tnh vi t l c cao, ngi ta s dng cht kch dc methyltestosterone vi thc n c hm lng m cao. mt s a phng cn nui trng qui m cng nghip lm thc n phc v cho vic nui mt s i tng nh ba ba, c chnh, Vic sn xut thc dng vin hn hp trong vi nm gn y phc v cho nui trng qui m cng nghip, m bo v thnh phn dinh dng, tan trong nc c m rng. nhiu cng ti lin doanh u t dy chuyn sn xut thc n phc v thy sn nh CP group ca Thi Lan, con c Ngy 24/3/1997 Nh nc ta ra quyt nh s 138 v tiu chun sn xut thc n cho i tng thy sn dng vin. Thnh phn thc n ny phi m bo cc cht dinh dng nh acid amin, vitamin, cc cht khong, Nhn dn ta cng bit ch bin cc loi thc n dng hn hp bng cc ngun nguyn liu sn c nh cm go, u nnh, c bt, nghu, s nui cho nhiu i tng c gi tr kinh t nh c ba sa, c tra, tm cng xanh, tm hm Chng hn, tm cng xanh cho n loi thc n gm c tp 199 kg, thc n cho g con c c 60 kg, u nnh 15 kg, hn hp vitamin v khong cht 0,5 kg. to mu sc cho sn phm sau thu hoch c mu sc ti, p, nhm nng cao gi tr sn phm, ngi ta cn cho n cc loi thc n c cha sc t. Theo Hip hi u tng M, cho tm s n theo 7 ch khu phn sc t v chia lm 3 thi k nh sau: astaxine nng 50, 100, 200 mg/g, -carotene 50, 100, 200 mg/g v to Dunaliella salina 100 mg/10 g khu phn thc n. 8.5. Cng ngh sinh hc trong phng v iu tr bnh cc i tng thy sn Trong nui trng thy sn, cc i tng nui thng hay nhim bnh v c khi b cht hng lot, lm thit hi rt ln cho ngi nui. khc phc tnh trng ny, trc y ngi ta dng cc loi thuc cho cc i tng thy sn. Ngi ta s dng cc thuc khng sinh nh: tetracycline, nhm sulfonamide, quilonone, erythromycine, (Nguyn Phc Tuyn, 2002), hoc oxytetracycline, sulpha c trn vo thc n, cng c th thm cc loi vitamin iu tr bnh, ng thi tng sc khng cc i tng nui. Chng hn, ngi ta s dng kt hp oxytetracycline + sulpha iu tr nhim Areomonas hydrophilla c m (b Thy sn, 1996) hoc ganamycine cha bnh en mang tm cng xanh. Theo Thi B H (1998), mt s thuc cha bnh tm c gii thiu v s dng c hiu qu trn mt s bnh. Tuy nhin, s dng cc loi khng sinh khi cha bnh lm cho cc i tng nui chm ln,
dng qu liu s gy d tha khng sinh trong sn phm thu hoch, lm gim cht lng v c th gy ng c cho ngi tiu dng. khc phc tnh trng ny, ngi ta dng gii php sn xut cc loi vaccine phng bnh, hoc ng dng cc qui trnh cng ngh sinh hc phng bnh. Gn y b Thy sn gii thiu ch phm VIRUTO phng bnh trng tm s v mt s loi tm khc rrt kt qu (theo VTV2).Yu Chen Chi (i Loan) pht trin mt loi vaccine kt hp bng nguyn l ca chui thc phm kt hp vi s ti t hp ca cng ngh. ng cy vi khun E. coli suy yu pht huy tc dng lng khng th c bit to ra phn ng min dch. Kt qu ny em li li ch cho ng dn nui c vng chu , c tnh c th t li nhun khong 3 t USD. Trong chn on pht hin bnh sm rt c ngha. Trong nui trng thy sn ngi ta ng dng cng ngh PCR l phn ng tng hp dy chuyn nh polymerase do Karl Mullius v cng s pht minh nm 1985 a vo chn on pht hin s nhim bnh, nht ;l cc loi thy sn trc khi a vo nui th. K thut ny cho php ti to theo mun mt mnh DNA khng theo php nhn dng v tnh, ngha l khng a mt on DNA vo c th sinh vt m ch s dng nhng mi chuyn bit. Nhng mi ny l nhng on DNA ngn, c kh nng bt cp vi mt u ca mch khun, trong vng mt pht to ra mt lng cc mch DNA. Hin nay nc ta, mt s a phng , cc vin nghin cu, cc trng i hc nhp cng ngh ng dng PCR ny vo qun l ngun ging v kim sot bnh. mt s a phng xy ra tnh trng tm ging th vo ao nui b cht hng lot do stress mi trng nui. Cng ti Long Sinh-Khnh Ha c hng dn mt s gii php nh trnh tng pH trong ao nui, iu chnh oxygene ha tan v trnh s dng qu liu khng sinh trong iu tr khi nui tm. Vaccine s dng phng bnh cho c nui c s dng ph bin. Cc loi bnh do vi khun gy ra c cng ti Th y Thy sn (Aqua healh Lid) ca Canada sn xut nh Lipogen triple v Lipogen forte cng vi Vibrogen-2, Furogen, Fry-vace, Ermogen, Renogen v Gaffogen. S dng cc loi vaccine ny c hiu qu rt r rt. Cc loi vaccine khc c sn xut bng cng ngh chuyn gene c gi l vaccine DNA. Mt chng trnh nghin cu ti Vin Locb Osawa (Canada) do tin s Heather Davis lnh o nhm nghin cu cho bit mt s th nghim trn c i vi hepatits c xem l mt m hnh chng minh tim nng ca loi vaccine ny trn cc loi c nui (Seffon Dixon, 2000). 8.6. Cng ngh nui trng trong x l mi trng nui Trong nui trng thy sn, trc khi ng ging hoc th ging nui, i hi phi x l mi trng, bao gm ao (b nui), ngun nc
cung cp vo v nc thi ra. i vi ao nui sau khi xong mt v nui, trc khi th ging mi, ngi ta tin hnh x l ao nui bng bin php th cng v dng mt s ha cht ty dit cc mm bnh. V d ao nui tm s, ngi ta x l ao nui bng mt trong hai phng php sau: x l t v x l kh. Tc l, ngm nc vo trong ao hoc phi kh mt ao, sau san bng nn y, xi nn y dit vi khun gy hi v lm cho cc kh c gy hi bay ln. Trc khi bm nc vo ao nui th ging, ngi ta cn dng vi bn nhm dit cc loi ch hi v iu chnh nng pH trong ao. Ngoi ra, trong vic qun l ngun nc nui, ngi ta cn dng mt ao lng x l bng zeolites hoc dolomite lm sch ngun nc v dit cc loi vi khun gy bnh. Theo Nguyn Vn Ton (chi nhnh Ven bin trung tm Nhit i Vit-Nga) th s dng h thng lc sinh hc phc v cc tri sn xut ging tm s v tm cng xanh Nha Trang, Khnh Ha mang hiu qu tt. Hin nay, cc ch phm sinh hc dng x l lp bn bn y ao cng c s dng rng ri. Cc ch phm ny c im chung l hn hp nhiu chng vi sinh vt hu ch v cc enzyme phn hy, chng c kh nng hn ch s pht trin ca cc vi khun gy bnh v dng cc lp bn lm ngun thc n. Nm 2000, cng ti Thng mi Dch v Mi trng Vit Nam nhp sn phm ES.2. AQUA KIT ca hng Enviroreps International (Hoa K). Ch phm ny bao gm nhiu chng vi sinh vt hu ch nh: Bacillus lichenformis, B. megatirium, B. subtilis, B. polymixa v Aspergillus oryzae cng nh cc enzyme nh: -amylase, Cc h sinh vt trong sn phm ny c kh nng cnh tranh vi cc vi khun c hi gy bnh, ng thi chng tit ra cc cht BDC dng t nhin c kh nng c ch sinh vt c hi trong ao nui. Cho n nay, trn th trng Vit Nam c n hng chc loi sn phm c dng tng t nh vy. V s dng cc m hnh nui nh nui xen canh: nui tm-sn xut mui, nui tm-rung la; hoc cc m hnh nui kt hp nh tm-s huyt, tm-rong sn, tm-c i cng c xem l hnh thc lm sch cc cht thi trong qu trnh nui. Gn y nht, nhm nghin cu Trng Vn Lung, V Tin Lng v mt s ngi khc dng rau cu cc (mt i tng c nhu cu v nhit v mn trng hp vi tm s) vi mt thch hp nui chung vi tm s. Khi quang hp, rau cu cc cung cp O2 cho tm s h hp lm gim bt sc kh cho tm. Rau cu cc li ht nhng thc n tha m tm khng s dng ht lm cho mi trng khi b nhim, s lng Coliforms (loi vi sinh vt gy bnh) gim. V vy, tm chng ln hn v khai thc c c hai i tng (tm s v rau cu cc) c gi tr kinh t cao, mt khc, li v din tch ao nui (khi
phi dng mt ao lng). Cng trnh ny c b Khoa hc v Cng ngh phi hp vi hng Sony (Nht Bn) tng gii Xanh Sony nm 2003. 8.7. Trin vng v ng dng cng ngh sinh hc trong nui trng thy sn - ng dng cng ngh sinh hc trong sn xut ging nhn to c cht lng cao, c h s ng ha thc n cao, to ra cc dng cht lng tt khng bnh bng phng php chuyn gene. - ng dng cng ngh sinh hc vo vic xy dng cc m hnh nui trng ph hp, kt hp vi vic lm sch mi trng nc thi, mang li hiu qu kinh t cao, u t thp. Sn xut vaccine trong phng bnh bng k thut ti t hp DNA hoc to dng n tnh. - ng dng cng ngh sinh hc sn xut cc loi protein n bo phc v ch bin thc n hn hp, sn xut cc loi thc n ti sng bng hnh thc t bo xc ca to, Artemia, Rotifer v.v. lm thc n phc v sn xut ging nhn to. - Phn lp v to ra cc hn hp vi sinh vt, cc enzyme s dng trong cc h thng lc sinh hc, x l nc thi v cc cht cn b t nui trng.
1. 2. 3. 4. 5. 6.
7.
TI LIU THAM KHO L Trn Bnh, L Th Mui, 1998.Cng ngh gene cy La.Hi ngh ton quc ln th nht v cng ngh sinh hc cy La ti Hu. Nxb Khoa hc v K thut H Ni. Trn Quc Dung. 2001. Nghin cu chuyn gene hormone sinh trng ngi vo c Chch (Misgurnus anguillicaudatus) bng vi tim. Lun n Tin s sinh hc, H Ni. i Duy Ban, L Th Cm Vn, 1994. Cng ngh gene v cng ngh sinh hc ng dng trong nng nghip hin i. Nxb Nng nghip H Ni. L Don Din, 1994. Cng ngh sau thu hoch trong nng nghip Vit Nam: Thc trng v trin vng. Nxb Nng nghip H Ni. Nguyn Mng Hng. 2004. Cng ngh t bo phi ng vt. Nxb i hc Quc gia H Ni, H Ni. ng Hu Lanh. Trn nh Min. Trn nh Trng. 1999. Cng ngh sinh hc vi cng tc ging vt nui, B. Trong: C s di truyn chn ging ng vt (ng Hu Lanh, ch bin), Nxb Gio dc H Ni, tr 433-469. Trng Vn Lung, 2004. Cng ngh sinh hc mt s loi To kinh t. Nxb Khoa hc v K thut H Ni.
8. L nh Lng, Quyn nh Thi. 2003. K thut di truyn v ng

dng. Nxb i hc Quc gia H Ni, H Ni. 9. Bernard R.Glick, Jackj Pasternak. 1994. Molecular BiotechnologyPrinciples and Application of Recombinant DNA. ASM Press. Washington D.C. 10. Chopra V.L, Anwan Nasin. 1990. Genetic Engineering and Biotechnology, Oxford and IBH Publishing CO.PVT, Ltd. 11. Dalton. D.C. 1982. An Introduction to Practical Animal Breeding. Granada Publishing Limited, Great Britain. 12. Peregrino G. Duran. 2002. Technical aspects of the recovery, handling and transfer of embryos. Copyright FFTC1970-2002. (http://www.fftc.agnet.org/library/article/tb151a.html). 13. Tp ch Th gii mi, Nxb Gio dc H Ni, S 613, tr: 39-42. 14. Clive James, 2004. Tm tt bo co s 32 ca ISAAA v tnh trng cy trng chuyn gene/cng ngh sinh hc c trng v mua bn trn th trng th gii trong nm 2004. Bn tin Cng ngh Sinh hc thng 01 nm 2005. www.biotechvn.com,vn
34 CNSH vi bo v sc khe cng ng
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2
CNG NGH SINH HC VI VIC BO V SC KHE CNG NG Chng III
Cng ngh sinh hc hin i trong cha bnh

Mc du sinh sau mun nhng nhng thnh tu m CNSH em li l rt ln. Nhng bc pht trin nhanh chng ca cc lnh vc nh: iu ch dc liu, to ra khng th n dng, to ra vaccine, thnh cng trong gii m b gene ngi... gp phn c lc cng vi con ngi trong cng cuc u tranh chng li bnh tt v khng ngng ci thin v nng cao cht lng cuc sng.
1. Sn xut dc cht v m phm v hi cho con ngi t sn phm t nhin v ti t hp gene trong sn xut vaccine
Trung Quc, mt t nc s dng nn y hc thc vt t 3.000 nm nay, khi xng nhng chng trnh nng cao dc in y hc c truyn ca mnh bng cch hng ti CNSH. c nhng chng trnh nghin cu tc dng sinh l v ha sinh ca cc hot cht thc vt trn ng vt th nghim; nhng chng trnh nhm vo cc lnh vc m hot cht c tc dng mnh nht. Theo Kuang An Kun, cu Gim c trng i hc Y s 2 Thng Hi, mt trong nhng ngi khi xng cho s hin i ha nn y hc c truyn Trung Hoa (Cn nhn mnh rng, hin nay Trung Quc, cc bc s sau 6 nm o to trng i hc Y, cn phi hc thm 5 nm na mi c cp bng chnh thc v ng y . Trong s 600.000 bc s ang hnh ngh, ch c 1/3 l c bng ny. Chng trnh o to chuyn khoa ko di ny khc hn chng trnh o to 1 nm vo u nhng nm 1960, ch cung cp nhng kin thc ti thiu cho hng nghn thy thuc chn t, lm cho h ch c th thc hin c nhng sn sc c bn nht, ghi n vi chc loi thuc v c ng cho phong tro v sinh vng nng thn Trung Quc). Nhng nghin cu gn y hn nhm vo vic phn lp t thc vt nhng hot cht, c th sn xut thuc vi s lng ln hn v s dng vi liu lng chnh xc hn. Bi v rng, thc vt thng sn xut
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cc hot cht vi s lng t i, hoc nu khng th ch t liu lng khng ging nhau trong sn xut. Trong cc vin Ha sinh v Ha hu c thuc vin Hn lm Khoa hc Trung Quc Thng Hi, ni m insulin c tng hp t nm 1966, ngi ta c gng chit xut cc loi dc liu nhm tng hp chng v tng hp c cc cht c cu trc t do c th c tc dng iu tr cn mnh hn. Mt trong cc v d l mt loi cy ph bin Trung Quc ni ting do tc dng gy sy thai, hot cht ca n c xc nh l mt protein c 230 acid amin. Protein tinh khit ny c hiu lc vi liu ch cn vi microgram. Hn 1 triu ph n s dng n. Ngi ta nghin cu b phn c hot tnh ca phn t protein ny v thm ch tng hp chng (Nature, vol. 318, No 6043, 21-23/11/1985). Qua ng dng CNSH, cc thc vt c mi m ng thi cng thng c tnh cht cha bnh (cc tinh du) c th tr thnh ngun nguyn liu tt cung cp cc cht cho vic sn xut dc cht. Artemisinine hay Quinghaosu (QHS) l mt loi thuc t nhiu th k dng chng st rt do cc nh khoa hc Trung Quc chit xut t cy ngi hoa vng mt nm (Artemisia annua). Ngoi quinine, mt alcaloid do nh khoa hc ngi Php l Pelltier chit xut nm 1934 t v cy Cinchona succirubra (Canhkina) v c dng cha bnh st rt cho n tn nhng nm 1950, khi thuc cha st rt tng hp ha hc ra i, nhiu thc vt khc cng c s dng vo mc ch ny, nh l Dichroa febrriusa (cy thng sn, c mc min Bc nc ta), mt loi thc vt Trung Quc. Alcaloid ca n l febrriugine. Tuy vy li qu c khng dng c cho con ngi. Nm 1967, theo quyt nh ca Chnh ph nc CHND Trung Hoa, nhm bt u chng trnh tng quan y cc cy thuc trong nc, ngi ta tp trung ch n Artemisia annua (cy ngi hoa vng mt nm) hay l cy Quinghao, thuc h Compositeae, trong bao gm c A. abisinthium (cy ngi trng) m cy l nhng nm 1920 vn dng trong sn xut ru apxin. Cy A. dracunculus (cy ngi gim) vn c bit n nh mt gia v v ch gim thm. Cy A. tridentata (cy ngi ba rng) min ty Hoa K, A. apiacea (cy thanh hao), A. capillaris (nhn trn Trung Quc) l nhng loi ngi dng trong dn gian cha st. QHS l mt sesquiterpene lactone c cha nhm feroxide v khc vi phn ln cc thuc chng st rt khc l khng cha d vng N. Cc dn xut mu ca QHS nh hydroquinghaosu artemether v sodium artesunate tc dng nhanh trn cc bnh nhn th no ang hng ti hn m; trong khi quinine hay chloroquinine tc dng rt chm th sodium arsunate lm khi hn m sau 12 gi.
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V m phm, cht mu thc phm v hi cho con ngi Trong thi gian qua, nhiu ca hng bn m phm v cht mu thc phm gy tai hi n sc khe ca con ngi cng khng t. B ngoi th qung co y l nhng cht mu thc phm t nhin hoc m phm c cha cc vitamin, cysteine, cc cht dit khun c trong l, hoa qu v.v. nhng thc cht dng cc cht mu hoc m phm bng cc cht ha hc. Cc cht mu ha hc c u im l mu ti hn, gi mu c lu hn v gi thnh r hn so vi cc cht mu t nhin. Nhng nhiu ngi dng cc m phm b d ng, l lot da, pht sinh nhiu bnh khc, a n nhng hu qu khn lng.Ngay Vit Nam nm 2000, c ti 45 ngi cht v 3800 ngi b ng c thc phm do dng cht mu thc phm bng ha cht. Nhng nm gn y, ngi ta i su vo vic nghin cu mt s cht c hot tnh sinh hc to v ng dng n. Mt trong nhng cht c hot tnh sinh hc c pht hin l phycobiliprotein. Ni chung, phn ln cc sc t thu nhn nh sng nm trong cc c quan t c bit, l phycobilisome. Phycobilisome c th tch chit c t t bo dng nguyn vn. mt s loi to nh to lam c nh N 2 khng c chloropyll b. Vai tr ca n trong quang hp c thc hin bi biliprotein. Cc protein ny c mu r nt nh cc nhm mang mu lin kt ha tr vi chui polypeptid. To v to lc thng cha c hai loi biliprotein l phycocyanin v phycoerythrin thng gi l dng C hay R (ty theo ngun gc ca chng l Cyanophyceae hay Rhodophyceae) Dng BR phycocyanin (B l to hng min Euchema) l mt biliprotein c nht mang c hai nhm mu ny. hu ht to lam u cha allophycocyanin. C-phycocyanin v C-phycoerythrin, tuy nhin mt s loi to lam khng pht hin thy phycoerythrin. (Donalt A., Bryant et al., 1976). Qua nhng nghin cu v quang ph hp th gip cho ta to iu kin tch, tinh sch cc nhm mu dng vo nhiu lnh vc khc nhau trong c vic sn xut m phm v cht mu thc phm. Hng Dainippon Chemical and Ink. Co. (Nht Bn) u t mt c s vt cht i tr to Spirulina platensis (18.000 m2) ti Thi Lan ch yu khai thc nguyn liu cho ch phm Linablue A. Ch phm ny cha 30% phycocyanin v c dng nh mt cht mu thc phm v m phm c ngun gc t nhin.Th trng hin ti trn th gii l 10 tn ch phm/nm vi gi bn 170 USD/kg, trong khi nhu cu l 200 tn/nm. Theo cc chuyn gia hng Cyanotech (M) th hng ny sn xut phycobiliprotein t Spirulina v Porphyridium trong nhiu nm nay v bn khong 40.000 USD ch phm (Report of Cyanotech Inc., 1985).
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Vin Du m Php, cng ti Soda-Texcoco ca Mexico l nhng nc sn xut Spirulina maxima xut khu. Nht Bn, M v cc nc chu u l nhng nc ch yu nhp khu bt Spirulina b sung ngun dinh dng cho thc phm v m phm. Khng nhng cc loi vi to m ngay c to ln cng c nui trng qui m rng. Sn phm ca n ngoi vic s dng trong cc nn cng nghip khc nhau, cn dng trong thc phm m phm nh nc gi u , kem bi mt, thuc nh mng tay, thuc xt tc, son bi mi, sp k lng my, v.v. Cc m phm sn xut bi cc nguyn liu t thc vt th rt an ton cho vic tiu dng ca con ngi. Bi v, n l nhng cht mu t nhin mc du trong qu trnh chit xut c nhiu chi ph tn km nn b t hn. Chng ta cn nghin cu ci tin gi thnh h v gi mu c lu hn v mu ti hn hin nay.
2. Nui cy m v t bo nhng cy thuc c gi tr cao

2.1. Sn xut khng sinh Nn cng nghip sn xut thuc khng sinh ra i t nhng nm 40 ca th k trc v c pht trin rt mnh v nhng l do sau: . Nhu cu s dng i vi thuc khng sinh rt ln v c xu th ngy cng gia tng; . Thuc khng sinh u tin l penicilline G (benzylpenicilline) ch khng c vi khun gy bnh gram dng nh Streptococcus v Straphylococcus, m thc t i hi phi c thuc khng sinh c cc vi S CH3 khun gy bnh gram m nh E. coli v Pseudomonas; . Thuc khng sinh hay gy d ng, cn CH c C CH3 khng -CH2CO NHCH - phi nhiu loi sinh khc nhau khc phc hin tng trn; N a OC CH COOH . Penicilline khng bn trong mi trng acid ca d dy, do b cn phi ci bin n v trn c s to ra nhng thuc khng sinh mi Benzylpenicilline c hiu lc v ph tc H H2N ng rng hn; Trong thc C phi lun to ra thuc khng sinh mi, v vi khun t ` S gy bnh ni chung CH2ln 2thuc v lm gim hiu CH2 ca thuc rt s CH CH2CONH CH CH lc HOOC nhanh. Th d: Straphylococcus lm quen penicilline kh nhanh, v n c OC N C-CH2OCOCH3 kh nng tng hp enzyme penicilinase (-lactamase) thy phn lin kt C amide ca vng lactam v lm mt hot tnh ca penicilline. (hnh III.1.). T nhng nm 1940 Cephalosporia sut toCOOH n nay, nng penicilline lun gia tng ch yu thng qua vic to cc chng Penicillium chrysogeneum t -CH-COCl bin (nh chiu x tia UV, tia X v s dng cht gy t bin (hnh III.3.) NH2 C C-CON C Ampicilline CH3 O Cloxacilline
Hnh IV.2. Benzylpenicilline v mt s penicilline bn tng hp a- V tr ct mch bn ca penicilline acid. b- V tr ct vng lactam

n v penicilline 100.000
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50.000 n v/ml
1.000 10
2n v
1940
/ml 1950
1960
1970
1980
S CH Hnh III.1. Qu trnh phn2 lp chng sn xut CH2CO NHCH CH C CH2 a OC N CH - COOH
b
Benzylpenicilline H2N HOOC H C S CH2CH2CH2CONHCHCH CH2 OC N C CH2OCOCH3 C Cephalosporine
-CH-COCl 2 USDA bang Illinois NH C C-CONRRL-1951-B25 N C UV Vin Cargenie v X-1612 trng CH2 Tng hp H Ampicilline O Cloxaccilille UV bang Minesota V Hnh III.2. Benzylpenicililline v mt s penicilline bn tng hp. Wis.Q-176 a-V UV tr ct mch bn ca penicilline acid. b- V tr ct Wis.B13-D10 vng lactam Trng H Tng hp Ngoi ra, ngi ta cng to c bang Wiscosin v cephalosporine streptomycine S Wis.47-638 l khng sinh khng vi khun gram m do vi khun dng si Actinomyces S v nm Cephalosporium to ra. Tuy Cc phng php l khng sinh thuc cephalosporine to t bin Wis.47-1564 S- Chn lc khi s nhm penicilline, nhng n khng gy d ng t nhin dng (Hnh III.2.) Ngoi ra, bngS cch thay th nhmUV- chiu x bng tia UV chc mch bn ca vng -lactam, Wis.48-701 nhiu loi khng sinh bn bng tia X c c tnh tc X- Chiu x tng hp ngi ta to ra c NM- x l bng methyl-bisng ht sc a NM Th d ampicilline l sn phm bn tng hp ca dng. Wis.40-133 ch l n cha thm nhm NH mch bn. (B-chloroethyl)-amin penicilline, ch khc 2 Wis.51-20 Tuy nhin, s thay i ny lm cho n c ph hot ng rt UV rng, thm ch cn khng c c mt s vi khun gram m gy bnh E-1 T E-1 n E-15 thng qua x l t bin bng methyl-bisCng ti Lilly (B-chloroethyl)-amin S Chng cui cng
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ng h hp (Haemophillus influenza), tiu ha (Schigella v Salmonella) v bi tit (E. coli Proteus). c bit do khng mt hot tnh trong mi trng acid ca d dy, nn ampicilline l loi thuc ung c, ch khng phi tim nh penicilline. Tng t nh cloxacilline cng khng phn hy trong d dy bi enzyme -lactamase, nn n thng c ch nh s dng km theo ampicilline cha tr cc bnh do Staphylococcus (vi khun gy bnh c kh nng to enzyme -lactamase) gy ra.
Penicillium chrysogenum NRRL-1951, Peoria, IL
Chng u tin Phng th nghim ca
E-15-1 Hnh III.3. To cc chng Penicillum chrysogeneum t bin nh chiu x tia UV, tia X v s dng cht gy t bin
2.2. Sn xut thuc bng cng ngh nui cy t bo Trong thc t rt nhiu cht c hat tnh sinh hc khc nhau (ch yu l cc cht th cp) thng s dng trong ngnh y t c to ra trong
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qu trnh nui cy t bo ng thc vt nguyn vn (vi b nhim sc th khng thay i). Chng c to thnh trong qu trnh nui cy m v t bo thc vt. Th d c th nht l s tng hp interferon trong qu trnh nui cy t bo nguyn bo bch huyt (lymphoblaste), l k thut to khng nguyn lm nguyn liu sn xut vaccine v to khng th n dng. Tt c chng u l nhng sn phm y hc cao cp v c tng lai rt to ln. 2.3.Sn xut thuc nh cc qu trnh bin i sinh hc c bit Bn cht ca n l cng ngh s dng vi sinh vt thc hin mt cng on kh khn quyt nh no trong chui bin i ha hc ca mt s cht c hot tnh sinh hc c s dng lm thuc cha bnh. Cc cng on ni trn v nguyn tc rt kh thc hin bng con ng bin i ha hc thun ty v rt tn km, Th d chng nm mc Rhizopus arrhizus thc hin c phn ng hydroxyl ha v tr C11 ca nhn progesterone l mt phn ng rt tn km v kh thc hin v mt ha hc. Bng bin php bin i sinh hc c bit ny, ngy nay ngi ta c kh nng sn xut c nhiu loi thuc cha bnh c ngun gc t steroid t nhin vi gi thnh r hn nhiu so vi trc kia.
Mt s thuc c ngun gc steroid Thuc steroid Cng dng Glucocorticoid Cortisone Bnh thiu mu, thiu huyt, c ch ty no Hydrocortisone Bnh tm thn addison Prednisone Bnh d ng, hen suyn, l lot cc b Dexammethasol Hormone steroid Thuc c ch min ch dng khi ghp c quan ca ngi Testosterone Estradiol Cha bnh khp v vim lot d dy Progesterone Cha tr bnh tm thn, bnh ri lon kinh nguyt, iu tr ung th v v chng r xng
3. Cng ngh sn xut sn phm y hc bng t bo ci tin

Bn cht ca vn ny l vic s dng cc t bo sng c ci bin (bng cch lai hoc thay i b my di truyn) sn xut ra cc sn phm c hiu cn thit cho i sng con ngi. Sau y chng ta s
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xem xt hai cng ngh in hnh nht: cng ngh sn xut khng th n dng v cng ngh sn xut thuc protein nh DNA ti t hp. 3.1. Cng ngh sn xut khng th n dng (monoclonal antibody) V bn cht khng th n dng l tp hp ca cc phn t khng th ng nht v mt cu trc v tnh cht, c to bi dng t bo lai gia t bo ung th myeloma vi t bo lymphocyte ca h min dch ng vt hoc ngi, trc khi xut hin cng ngh s dng t bo lai sn xut khng th. Vic to ra mt lng ln khng th c tnh cht xc nh no l mt cng vic cc k kh khn. L do ch yu l v cc phn t khng nguyn (antigene) kch thch t bo ng vt c xng sng to ra nhiu phn t khng th (antibody) khc nhau. Vic tch ring cc phn t khng th ny l mt vic rt kh khn v tn km.
Khng nguyn PEG
T bo myeloma khng khng HAT T bo gan khng HAT Trn Nui cy trn mi trng cha HAT
Hnh IV. 4. S cng ngh to khng th n dng Hnh III.4. S cng ngh to khng th n dng
Do , cng trnh nghin cu hon thin cng ngh to khng th n dng ca Milstein v Kohler l mt bc ngot trong lnh vc ny. Cng ngh ny cho php m ra mt lot nhng ng dng quan trng trong
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y t cng nh trong nghin cu v sn xut. Ni dung ca cng ngh ny c m t trong hnh IV.4. bao gm cc bc sau: - Tim khng nguyn vo chut. Tch t bo gan chut (l ni khng th c to thnh). Trn t bo gan chut (khng c HAT) vi t bo ung th myeloma (khng khng c HAT). B sung PEG (polyethylen glycol) vo dch huyn ph cha hn hp hai loi t bo trn. - Nui hn hp t bo trn mi trng chn lc cha HAT (hn hp ca hypoxanthine, aminopterine v thymidine). T bo gan chut s pht trin bnh thng trong mt thi gian, cn t bo myeloma do b khuyt HAT, nn cht ngay. - Hn hp nui cy to ra t bo lai. Kt qu l trong hn hp t bo ni trn, t bo myeloma cht ngay, t bo gan cht sau hai tun nui cy, cn t bo lai pht trin bnh thng, v n c hai c im: phn chia v hn (nhn c t t bo myeloma) v cc cht trao i cn thit (do t bo gan chut cung cp). 3.2. Sn xut thuc nh cng ngh s dng DNA ti t hp Bn cht ca cng ngh ny l thay i b my di truyn ca t bo bng cch a gene l vo v bt n hot ng. C ngha l, bt n to ra mt lng protein hoc peptide cn cho con ngi, m trc bn thn t bo khng tng hp c. C th ni, cng ngh ny to ra bc ngoc ln lao trong lnh vc sinh hc. Cc sn phm do n to ra kh a dng v mt s hin ang c sn xut qui m cng nghip. Di y l mt s v d: + Sn xut insulin bng cng ngh s dng DNA ti t hp. Hin nay khong 1-2% dn s cc nc pht trin b bnh tiu ng. cha tr ngi ta phi s dng insulin, l protein cha 51 amino acid c tng hp tuyn ty ca b v ln. Tuy nhin, v cu to ca chng hi khc vi insulin ca ngi nn trong mu ca bnh nhn iu tr insulin b bao gi cng xut hin khng th i vi insulin b. iu ny gy ra mt hu qu khng mong mun lm mt mt phn hot tnh ca insulin v gim thi gian tc ng ca thuc. Vo u nhng nm 80 ca th k trc, hng Eli Lilly v Genetech Inc. ca M tm ra qui trnh sn xut insulin ngi bng cng ngh DNA ti t hp. l protein u tin to c bng cng ngh gene v nh cng ngh ny m hin nay insulin c sn xut i tr vi gi thnh r hn nhiu so vi trc kia (hnh III.5.) + Sn xut interferon bng cng ngh s dng DNA ti t hp. Interferon l nhm protein cha khong 146-166 amino acid. Chng xut hin trong t bo ng vt c xng sng khi b nhim virus v gip t bo khng li virus.
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Ngoi interferon cn c hai c tnh rt gi tr: mt l, chng c ch s sinh sn t bo (nn c s dng cha tr bnh ung th); hai l, chng gia tng hot h min dch ca ngi v ng vt. C ba loi interferon: -interferon, -interferon v interfefron, khc nhau bi s amino acid v s th hin hot tnh.
Tng hp gene m ha si A insulin, khu vo gene m ha galactosidase ca plasmid chuyn chuyn vo E. col Tng hp gene m ha si B insulin, khu vo gene m ha galactosidase ca plasmid chuyn plasmid vo E. coli
Gene si A Gene si
Gene Plasmid si A Plasmid Si A E. coli -galactosidase Plasmid Si B E. coli -galactosidase Gene si B
Nui E. coli trn mi trng c galactosidase nu enzyme -galactosidase v cng vi n si A v B ca insulin cng c tng hp. Lm tan t bo x l BrCN tch si insulin ra khi -galactosidase Si A ca insulin Si B ca insulin
Oxyha S-S Phn t insulin ngi S-S Hnh III. 5. Cng ngh sn xut insulin bng DNA ti t hp
Cho n thi gian gn y, interferon ch yu c to ra t t bo bch cu hay nguyn bo bch huyt ca ngi b nhim virus. Do vy, gi thnh rt t v rt him (khong 40-50 USD/1 triu n v vo nhng nm 1980), m thng thng trong iu tr ung th mi ln phi tim cho bnh nhn ti 3-6 triu n v v phi tim nhiu ln. Khong hn 10 nm tr li y, ngi ta to ra c cc gene m ha tng hp ba loi
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interferon ni trn v xy dng c qui trnh cng ngh sn xut interferon trong t bo E. coli v nm men. Nh vy h gi thnh ng k ca interferon, hin nay khong 3-5 USD/1 triu n v (1994). Vin Sinh hc Nhit i thnh ph H Ch Minh t nm 1989 hp tc vi cc nh khoa hc thuc vin Hn lm Khoa hc Ucraina trin khai cng ngh sn xut interfefon ni trn thnh ph H Ch Minh. Bc u h to c hng chc t n v interferon, s thuc ny c th nghim c kt qu trn ngi bnh bnh vin Ch Ry thnh ph H Ch Minh. Hi vng trong mt tng lai gn thuc interferon s l sn phm gn gi v hp ti tin i vi ngi Vit Nam. Tng t nh interferon, mt sn phm khc gip c th tng kh nng min dch l interleukin cng c cc nh sn xut theo cng ngh s dng DNA ti t hp. Hin interleukin v interferon ang c th nghim cng vi mt s bit dc khc cha tr bnh ung th v bnh AIDS. + Sn xut hormone pht trin ca ngi nh DNA ti t hp. Hormone pht trin ca ngi l protein cha 191 amino acid tng t vi MW khong 22.000 dalton. Thiu n c th con ngi s b ln. cha tr, ngi ta phi tim hormone pht trin cho nhng a tr thiu hormone ni trn. Trc y, hormone pht trin c tch t tuyn h no ca ngi cht. Tuy nhin, phng php ny cn mt s hn ch. Hin nay, hormone pht trin c sn xut bng cng ngh s dng DNA ti t hp thng qua E. coli. + Sn xut vaccine bng cng ngh DNA ti t hp. Cng trnh u tin ca Louis Pasteur (thng 7/1885) m u cho k nguyn u tranh chng cc bnh nhim trng v phng chng bng tim chng. Cho n thi gian gn y, ngi ta vn s dng vaccine sng (l vi khun v virus gy bnh b x l lm yu i) lm khng nguyn kch thch to khng th cn thit trong c th ngi v vt nui. Do vy, xut hin nhu cu to vaccine th h mi trnh c nhng hn ch ni trn. Cho n nay, bng cng ngh s dng DNA ti t hp ngi ta sn xut c protein v ca mt s virus nh: virus bnh l mm long mng, virus bnh di v vim gan B. Trong s ny, vaccine u tin l vaccine vim gan B ch t protein v ca virus vim gan B (l bnh ph bin ngi Vit Nam). Vaccine ni trn (c bn cht l protein) c tng hp v tch ly trong t bo nm men hay t bo ng vt cha gene m ha protein hoc peptide n c sn xut bng cng ngh DNA ti t hp.
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ng dng enzyme trong y hc. Trong mt tng lai gn, enzyme s c ng dng rt rng ri trong ngnh y t lm u d cho cc thit b y t v cha bnh. Hin nay, mt s enzyme nh glucosoxydase, hexokinase, cholesteroloxydase, esterase, urease, alcohol dehydrogenase, v.v. c s dng kh rng ri trong ngnh y t. in hnh nht l vic s dng glucosoxydase c nh trn b mt in cc platinum trong thit b phn tch hm lng glucose trong mu. Bn cht hot ng ca in cc nh sau: Enzyme glucosoxydase c gn trn mng in cc, khi in cc tip xc vi dch mu cha glucose th n s xc tc phn ng: Glucose + O2 Gluconic acid + H2O2 Do , nu trong mu cng nhiu glucose th cng nhiu oxygene t do c ghi nhn trn in cc platinum. Qua , ngi ta s xc nh c hm lng glucose ca mu phn tch (th d nh trong mu chng hn) (hnh IV.6)
Glucose Mng gn glucosoxydase O2 H2O2 Gluconic acid Mng in cc
in cc platinum (cathode)
Hnh III.6. in cc gn enzyme glucosoxydase Trong y hc, enzyme cn c s dng loi b nhng cc tht m d tha gy nguy him cho mt s b phn c th ngi. Th d, ngi ta dng enzyme streptokinase v urokinase lm tan mu ng lm nghn mch mu. Hoc, enzyme cn s dng cha tr bnh lin quan n s thiu ht mt s enzyme trong ngi. u nm 1993, mt bc s ngi Italia cng tin hnh phu thut gene cho mt chu trai 4 tui o Sicile (Italia) b bnh di truyn bm sinh thiu enzyme adenosine desaminase ch v lm ln mt gene trong khong 40.000 gene, lm cho h min dch ca ngi cc k ri lon. Chnh enzyme ny ph hy mt trong cc sn phm trung gian sn sinh ra trong qu trnh trao i cht. V hi i vi t bo khc nhng li rt c i vi lyphocyte T (t bo bch huyt T). Khng c n h thng min dch dn dn b hng (ging nh
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bnh AIDS). Trc y, bnh ny v phng cu cha. Gn y, ngi bc s tin hnh ly mu ca ty xng ca chu ni sinh ra lymphocyte T, a virus v hi c mang gene ca enzyme ni trn vo mu ty , ri li a ngc tr li vo c th chu b. Liu php lp li 3 ln, mc enzyme trong mu tng. Chu b thy khc hn ln. Cng ngh gene tin hnh khng phi ln u tin. M c mt chu b gi ang sng bnh thng sau khi ghp gene t ngy 14 thng 9 nm 1990. Ba nh bc hc ngi M Frech Andreson, Michael Blaese v Ken Calver vin Sc khe Quc gia tin hnh chuyn gene enzyme adenosine desaminase cho mt chu b gi 4 tui mc bnh suy gim min dch phi hp ni trn (SCID) phi sng trong mt cn bung v trng. Thng 5/1993, nhn mt cuc hp bo, ng Ken Calver - ngi lnh o nhm chuyn gene ny ni: Tm thi chng ti hi lng vi kt qu thu c. Mt chu b hu nh hon ton trnh c nhim trng, nng enzyme t gn mc bnh thng. H min dch ca chu khe ln. Chu c th chy chi ngoi ph, i n lp hc, thm ch chi th thao nh cc chu khc.
4. S dng phng php di truyn phn t v k thut DNA ti t hp chn on v cha tr bnh
4.1. Chn on bnh di truyn thai nhi
T bo hng cu li lim T bo hng cu bnh thng
Tch DNA v ct bng nuclease on DNA cha gene m ha hemoglobin
Tch DNA m-RNA ca globulin Sao chp ngc +P*
Lai cc on DNA vi nhau Pht hin on DNA c hiu ca globulin bng phng php ng v nh du
Hnh III.7. Phng php ct phn on a hnh DNA
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Trc y, pht hin bnh ny thai nhi ngi ta phi ly mu th trc tip t thai nhi. iu ny nhiu khi gy tn hi cho thai nhi. Gn y, ngi ta bt u pht hin vn s dng phng php di truyn v k thut DNA ti t hp chn on v cha tr bnh ngi bt u c quan tm t u thp k 80 ca th k trc. Ln u tin k thut ny c p dng chn on xem thai nhi c b bnh thiu mu do hng cu li lim hay khng. Hin nay bnh ny bng phng php ct phn on a hnh RFLP (Restriction Fragment Length Polymorphism) DNA ca t bo dch xung quanh thai nhi. Sau so snh chng vi cc phn on DNA b tr tch t t bo hng cu bnh thng pht hin s c mt ca gene b hng. Phng php ny an ton v khng gy nguy him cho thai nhi. (Hnh III.7). Ni dung ca phng php RFLP nh sau: tch DNA t t bo hng cu ca c b v m c con b thiu mu do hng cu li lim, t ngi khe mnh v t dch bo quanh thai nhi tip theo ca ngi m ni trn. Dng enzyme endonuclease ct cc DNA ni trn thnh cc phn on khc nhau v tch chng bng in di. Mt khc, t t bo hng cu li tch m-RNA, cho tng hp ngc to DNA b tr cDNA (complementary DNA) gn 32p. Trn cc on DNA vi cDNA nhm to cc on lai ny (on ha hp c vi th t nucleotid ca cDNA) l on cha gene m ha tng hp globulin. ngi khe mnh gene glubolin nm phn on c di 7.6 kb. a b b thiu mu do hng cu li lim, gene ni trn nm phn on c di 13,0 kb, cn b m v thai nhi pht hin thy gene ny c mt c on 7,6 kb v 13,0 kb. iu ny chng t c ba u dng d hp t v mang gene bnh thiu mu do hng cu li lim. 4.2. Vai tr ca cc tc nhn di truyn trong bnh l hc Cc tc nhn di truyn trong mt s trng hp l nguyn nhn gy ra mt s bnh (trong c nhng bnh him ngho). C ch phn t ca s th hin bnh di truyn hin cn cha bit c nhiu. n c th d in hnh nht l bnh tiu ng do di truyn. N th hin mt s dng khc nhau. Nhm th nht bao gm cc bnh nhn cn tr, h t bo chu trch nhim tng hp insulin hu nh b cht hon ton. Do vy, c th ca cc bnh nhn ny khng t tng hp c insulin. Nhm th hai bao gm cc bnh nhn ln tui, h insulin c tng hp bnh thng. Tuy nhin, c ch hp th insulin li khng hot ng, nn c th vn lun lun thiu insulin. Nh s phn tch RFLP ngi ta pht hin thy c s khc nhau v s lng v th t sp xp gc nucleotid on gn vi u 5 ca gene m ha tng hp insulin. Bng phng php trn, ngi ta cng pht hin thy lin quan ca cc tc nhn di truyn vi s
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th hin ri lon teo c Duchenne v bnh tm thn tr tu km pht trin Huntington. Trin vng ca vic cha tr cc bnh c lin quan n di truyn rt ln. Trong vi nm tr li y xut hin mt s cch tip cn gii quyt vn ny. l vic gn b sung gene bnh thng vo b my di truyn ca ngi bnh hoc lai v cy t bo khe mnh vo c th ngi bnh cha tr mt s bnh him ngho v bnh di truyn. Tuy cn phi khc phc nhiu tr ngi, nhng cc bin php cha tr ny rt c trin vng trong tng lai.
5. Khng th n dng (monoclonal antibody)

Hybridoma v khng th n dng. Cc khng th n dng t t bo ung th: khi ngi ta tim vo chut hay th mt cht l, lp tc c th phn ng to ra mt loi phn ng c hiu chng li mt khng nguyn. Mt khng th c tnh c hiu cao v ch chng li mt khng nguyn. Trong c th ch c mt loi t bo c bit sn xut ra mt khng th . Nu ta tch mt m nui v chn ng dng cho ra khng nguyn m ta cn th ngi ta c th c mt dng to ra khng th c hiu vi loi khng nguyn. Nhng t bo m ta tch l t bo bnh thng nn sinh sn c hn (ch phn chia mt s ln ri cht). Trong khi , dng t bo ung th l bt t. Nh trn cp n, nm 1975, Kohler Milsstein pht minh ra c khng th n dng. Nguyn l ca n l: + Gy ra nhng t bo bnh thng c kh nng to khng th. + Lai nhng t bo ny vi t bo myeloma (t bo ty) ung th. Qu trnh ny c hiu qu khi dng virus Sendaii bng phng php dung hp t bo. + Chn cc loi m ta cn, l dng lai va mang tnh cht to khng th, va c tnh cht sinh sn v hn (gi l dng hybrodoma). Chng c ng dng trong nhiu lnh vc: - Tng nhy ca php th dng cc phn ng min nhim cc khng nguyn khng dung np (histocompability) th cc khng nguyn ca nhm mu, ca tinh trng, th c progesterol khng, th cc yu t lm ng mu. - Chn on xem mt bnh no c truyn theo ng sinh dc hay khng hay l vic chn on ung th. - V liu php, chnh n li thuc qu liu, gim nguy him trong trng hp ghp ty sng, cha tr ung th ng ch , ng v tr cn thit. - Dng lm tinh sch cc sn phm, nht l sn phm protein. Bng k thut di truyn ta c th nui cy vi sinh vt sn xut ra insulin.
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- Trong nghin cu gip ngi ta pht hin ra v tr ca protein. - Vic s dng cc khng th n dng s nhanh chng thay th cc phng php min dch v huyt thanh hc thng thng trong cc xt nghim sau: Th nht l xc nh mc hormone nh gi chc nng tuyn ni tit hoc thay i tng hp hormone do khi u. Th hai l pht hin mt s protein c ngha chun on khi u hoc mt s iu kin c bit trc khi sinh . Th ba l xc nh vi khun gy bnh. Pht hin cc thuc b cm trong mu hoc kim tra nng thuc trong mu v t chc nhm m bo liu dng thuc khng vt qu ngng gy c. Cc ng dng khc bao gm: . c ch phn ng thi loi khi ghp cc c quan. . Min dch ha th ng chng li cc khng nguyn tham gia vo sinh sn (chng th thai bng phng php min dch). . nh v khi u vi nhng khng th c hiu. . Tc ng c t nh hng ti t bo ung th, bng c t ha hc gn vi khng th. Cc cht sau y c th dng khng th n dng pht hin: vitamin, thuc gim au, thuc ng, khng sinh, thuc chng vim, thuc chng tn sn (chng ung th), thuc tim mch, thuc gy o gic, thuc h ng huyt, cc hormone ng vt v thc vt, thuc lm du v an thn , thuc li tiu, steroid (androgene, estrogene, progestin), thuc tr su, c t (aflatoxin). Nh tnh c hiu v chnh xc cao, s dng d dng, khng th n dng to nn mt nhnh pht trin mau l nht ca CNSH. Theo tp ch High Technology magazine" th trng khng th n dng t 15 triu USD nm 1982 n nm 1992 vt qu 5 t USD. Loi sn phm khng th n dng dng ch yu l cc thuc th nghin cu, thuc th cho chn on in vitro v cc khng th dng phn lp cc cht sinh hc. Khng th n dng c cc nh khoa hc Nht Bn s dng trong chn on ung th ng tiu ha, c cng ti CNSH Centocor Inc. Hoa K dng chn on ung th ty, trng i hc Y khoa Nht Bn, trung tm Ung th Quc gia v cng ti Kureha Chemical Industries dng chn on ung th tin lit tuyn. Khng th n dng c dng chn on v th nghim iu tr carcinoma gan nguyn pht.
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i hc California v trung tm Y hc Albert Einstein (Philadelphia) dng iode phng x gn vo khng th n dng sau cng ph c hiu cc t bo ung th gan. Khng th n dng chng khng nguyn c hiu ca lymphocyte T c ng dng quan trng trong d phng phn ng thi ghp khi truyn ty. Ngy nay cng vi CNSH sn xut cytokin (ging nh hormone tham gia iu ha cht ch gia tr li min dch dch th v min dch trung gian t bo), ngi ta cng sn xut cc khng th n dng dng trong chn on v iu tr min dch. Cc khng th n dng ca chut tuy c dng trong iu tr nhng c mt s nhc im. Mt s nghin cu so snh v tn sut lai to gia t bo lymphocyte ngi vi cc dng t bo khc v theo di tnh bn ca chng. Myeloma (t bo ung th c dn ra t cc lymphocyte bi tit khng th) ca chut NS-I) hp nht vi tn sut cao nht nhng km bn hn l cc t bo lai gia ngi vi ngi. Vn tm kim t bo thnh vin thch hp hp nht vi lymphocyte ngi l mt hng mi. Hin nay cn gp mt s kh khn trong vic tm kim t bo thnh vin thch hp. Ngi ta la chn mi trng thch hp trong lai to l hypoxanthine, aminopterine v thymidine (HAT) nhng t bo lai gia myeloma v lymphocyte c kh nng s dng hypoxanthine cung cp trong mi trng HAT. Trong cc phng php phn tch min dch phng x RIA (Radio Immuno Assay), RAST (Radio Allergo Sorbent Test) v RIST (Radio Immuno Sorbent Test), ngi ta xc nh hm lng khng nguyn bng cc o phng x ca phc khng nguyn khng th. Cn trong phng php ELISA (Enzyme Linked Immuno Sorbent Test), ngi ta xc nh hm lng khng nguyn bng cch o phng x ca phc khng nguyn khng th to ra enzyme c hiu gn trn n. Sau y l phng php RIA trong nhng phng php c nhy cao nht (hnh IV.8). Ni dung ca n nh sau: Gn khng nguyn ln ba th nhy cm vi n. Ra, loi b nhng phn t khng nguyn khng gn. B sung khng th n dng gn c hiu vi khng nguyn. Ra, loi b nhng phn t khng th khng gn. Gn cu ni c nh du ng v phng x ln khng th. Ra, loi b nhng cu ni khng gn. m ng v phng x bit s lng khng nguyn.
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Cc khng nguyn Ra Gn khng th Ra Gn cu ni Ra m phng x
Khng nguyn Khng th * Cu ni nh du * *
Hnh III. 8. Phng php A phn tch min dch phng x RIA Cc phng php khc ch khc phn ui gn vo khng th. Th d trong phng php RIA, RAST v RIST - phn ui gn c cha ng v phng x. Cn trong phng php ELISA - phn ui gn cha enzyme xc tc phn ng to mu. Ngha l, thng qua phn ng to mu bit c c (hay khng c) khng nguyn trong mu th. Hin nay cc phng php phn tch s dng khng th n dng ang nhanh chng thay th cc phng php phn tch min dch v huyt thanh truyn thng trong cc phn tch.
6. Cc vaccine v vaccine a tr
6.1. Cc vaccine Vaccine l ch phm khng nguyn gy trng thi min dch m khng gy bnh. Vaccine dng kch thch p ng min dch nguyn pht, lm tng t bo nh v kh nng p ng min dch nh khi tip xc vi khng nguyn ln sau. T 6 n 16 thng 7- 1885, ln u tin tm cch tim chng chng bnh di (mt bnh c kh nng t vong 100%) cho mt em b 9 tui b ch di cn, Louis Pasteur cng vi cng s ca mnh l bc s iu tr Grancher phi cy truyn 13 ln virus di trong ty sng ca th vi c tnh tng dn. Ngy 27- 7 -1885, em b c cu sng v khe mnh. Ngy 14-10-1888, vin Pasteur c thnh lp, Grancher trnh by phng php ca Pasteur v s liu thng k chng t hiu qu ca n: 5374 ngi c iu tr trong thi gian t 16 - 7-1885 n 01- 071888, t l t vong l di 1%. Nh vy, Pasteur m u cho k nguyn u tranh chng cc bnh nhim trng v phng chng bng tim chng.
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Vaccine sng hay vaccine gim c lc (Attenuated vaccines): l nhng chng vi khun c gim hot lc v c th sinh sn m khng gy bnh, nhng kch thch c sinh tng hp khng th bo v. Vaccine sng i khi khng n nh v c th quay tr li dng c gy ra bnh m l ra n phi bo v c th chng li. V d nh vaccine khng bi lit. Vaccine ny dng nh khng cn ph hp cho cc nc ang pht trin. Hn th na virus gim hot lc sau khi tri qua nhiu chu k nhn ln trong c th tr em c tim chng vn c th phc hi li hot lc gy ra cc triu chng bi lit. S liu thng k Hoa K chng t rng trong s 10 tr em c tim chng th vaccine gy ra 9 bi lit (vim ty xm) cho 3 trng hp. Virus vaccine cng c th gy ra 3 n 5 trng hp vim no trong 1 triu ngi c tim chng u ma. Vaccine bt hot: l nhng virus hay t bo vi khun nguyn vn em bt hot, l nhng protein phn lp t virus, thng gi l nhng vaccine tiu n v" (vaccine cm ch t haemagglutinin, vaccine chng vim gan B ch t khng nguyn b mt vim gan B HbsAg), l nhng c t vi khun bt hot (gii c t bch cu v un vn); nhng khng nguyn b mt tinh ch (vaccine vim mng no). S dng nhng vaccine bt hot th khng c kh nng phc hi li dng c v tnh lnh c kim tra tt hn Tuy nhin nhng vaccine bt hot li t hn v tnh min dch km hn so vi vaccine sng. Vaccine sn xut bng k thut gene: C th dng t bo vi khun hay nm men thay i bng k thut gene sn xut cc protein min dch vi s lng ln v dng ch nhng vaccine hiu qu. u im ca vaccine ny l: + Khng nguyn dng kch thch min dch c phn lp t phn lnh tnh, khng gy bnh ca vi sinh vt gy bnh, v c tng hp bng cc t bo vi sinh vt hay ng vt c lp rp gene, m bo c u im ln l tnh an ton ca qu trnh sn xut. + Sn xut vaccine bng vi khun c lp rp gene lm gim mnh gi thnh v thay th c cc cng on t tin bao gm mi trng nui cy m ng vt hoc phi bng cc mi trng nui cy vi sinh vt thng thng, tng i n gin. + Gi thnh cn gim thm na nh khng phi trang b tn km cho m bo an ton cao (v d vaccine thng dng chng bnh l mm long mng thng c gi cao do qu trnh sn xut i hi nhng nh xng c an ton cao). + Trnh c vic phi th nghim tnh an ton trn qui m ln, v vaccine khng cha tc nhn gy bnh.
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+ Gi thnh bo qun v vn chuyn cng h thp nh gim c cc yu cu v lm lnh hoc ng kh. Bng k thut gene ngi ta cng sn xut c cc protein v ca virus l mm long mng, di, vim gan B, herpes v cm. Vaccine k thut gene u tin cho ngi l vaccine vim gan B, ch t khng nguyn b mt (HBsAg), tng hp trong t bo nm men hay ng vt nui cy lp rp gene. Ch phm c tinh ch, loi b cc protein v on DNA ca chnh t bo ch. Mt v d khc l vaccine si kiu mi c ch Trung tm Nghin cu Vi sinh hc ng dng (Porton Down nc Anh), vi s hp tc ca i hc N hong (Belfast), u vit hn hn loi vaccine si thng dung ch t virus gim c lc. Vaccine si kiu mi cha hai thnh phn khng nguyn: mt ngng kt t hng cu v mt protein lin kt, c hai u c tng hp bng k thut DNA ti t hp. 6.2. Vaccine a tr Ngi ta c th ch cc vaccine a tr bng cch ghp ni cc gene thnh ra gene m ha cho protein lai ghp, mang c tnh min dch ca cc khng nguyn khc nhau. Valenzuela v cc cng s ca mnh, nm 1985 ghp on DNA m ha cho on glycoprotein D (gm 300 gc aminoacid) ca virus herpes vo on tin-S, m ha cho protein v gia ca virus vim gan B. Gene mi ti t hp ny c ghp vo t bo nm men. Khi hot ng, n tng hp cc ht virus kiu mi, mang c tnh khng nguyn lai v khng tc dng vi cc khng th khng HbsAg (vim gan B) ln glycoprotein D (herpes). Hot tnh khng nguyn khng c hai loi virus ny c gi li, tuy nhin cc lot virus ny khng gy ra c sinh tng hp khng th khng hai virus ny. Nm 1987, cc nh nghin cu vin Pasteur Paris (phn vin ng dng Cng ngh Di truyn v phn vin Nghin cu Vi khun ng rut) cng b tng hp c c t bch hu lai mang c hai tin khng nguyn b mt virus vim gan B (HbsAg). Khi tim chng vo ng vt th nghim th c t ny gy c s tng hp khng th khng c c t ln HbsAg. Nh vy, u im ln ca vaccine a tr l c th cng lc s dng nhiu khng nguyn trong c th nhng vacine n tr li c ng dng mnh m hn bi tc dng c tr ca n. Vi s pht trin mnh m ca ln sng cng ngh sinh hc trn th gii, va qua, cc nh khoa hc Cuba va cng b sn xut thnh cng vaccine nhn to phng chng vim phi v vim mng no. Khc vi cc dng vaccine c sn xut theo phng php truyn thng, loi vaccine
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mi sinh khng th c to ra bng cch tng hp cc cht ha hc. Loi vaccine mi ny cha c cng b gi chnh thc nhng chc chn gi s r hn loi vaccine c vn c gi n 3 la mt liu. Chnh ph Cuba d kin s tim vaccine mi cho tt c cc tr em di 15 tui ca nc ny. Cng vi Cuba, Trung Quc cng cng b iu ch c vaccine phng chng hi chng vim ng h hp cp (SARS). Vic th nghim vaccine trn Kh thu c cc kt qu kh quan v hon ton khng gy ra phn ng ph. D kin n cui thng 04 -2004 ny s c 1.500 liu vaccine chng SARS c xut xng v sau s c thm 20.000 liu na c sn xut. Hng Vax Geneem gn ti ch trong vic n lc iu ch vaccine phng chng AIDS. Nhng nghin cu ban u cho thy nhng kt qu y ha hn. Song song vi nhng nghin cu ca Vax Geneem, cc nh nghin cu Anh th khong trn 20 loi vaccine trn ngi v ng vt. Gio s Andrew Mc Michael thuc phng th nghim Oxford bo co v mt trong s ti hi ngh khoa hc Lucy Dorrel: l vaccine c iu ch t DNA; on ny khi a vo c th th c kh nng m ha protein ca virus HIV, loi protein gy nn nhng phn ng t v mnh nht ca c th, theo hai cch: hoc di dng mt DNA trn hoc cng vi mt virus gy bnh u ma b suy yu. Nhng nu tim c hai cng mt lc l tt nht. Anh v c chu Phi, cc nh khoa hc th nhng mi tim u tin trn ngi v ang ch i kt qu. Trong thng 09/2004, h tim t hp vaccine trn cho nhng ngi b nhim HIV trn vi hy vng rng t hp s kch hot h khng v cng vi liu php phi hp ba loi thuc hin ang s dng s gip con ngi chin thng c bnh AIDS. Ln sng cng ngh sinh hc cng tht mnh m ti khu vc chu -Thi Bnh Dng khi m Vit Nam cng cng b sn xut thnh cng hng trm ngn liu vaccine thng hn vipolyssaccharide t tiu chun ca t chc y t th gii v hiu lc v an ton, vi gi ch bng 1/7 so vi gi nhp t Php, p ng nhu cu phng dch v xut khu. Vit Nam cng rt t ho khi m va qua dch st gia cm ang lan trn khp chu . Trong khi vin Pasteur Nha Trang, vin Sinh hc Nhit i thnh ph H Ch Minh cng b sn xut c vaccine chng bnh st gia cm khi gii m hon ton b gene ca virus st gia cm (H5N1). (theo tin tc VTV). Gn y nht (thng 12/2005), Tin s Cao Bo Vn, vin Pasteur thnh ph H Ch Minh gii m c virus H5N1 cc loi gia cm khc nhau. Gio S Nguyn Thu Vn vin Pasteur H Ni cng cng b sn xut c vaccine A H5N1 trn t bo
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thn kh tin pht, chng bnh dch do gia cm ly qua ngi, ang ch xc nhn ca Quc t sn xut hng lot vo u nm 2006.
Hnh III. 9. Virus H5N1
Mt thnh tu cng va mi c cng b mi y l chng ta sn xut c khng nguyn khng nc c rn. Tuy mi bc u sn xut liu lng cn hn ch, gi thnh cao.Tuy nhin trong tng lai s c sn xut vi lng ln nhm p ng y nhu cu ca con ngi v loi vaccine ny. Mt hng mi trong ch to vaccine l pht trin cc th khng th (antidio type antibodies).Khi khng th c sinh ra chng li mt khng nguyn th mt loi khng th th hai khng li khng th th nht cng s xut hin. Khng sinh Nm 1929, Alexandre Fleming tm ra tc dng ca khng sinh khi nghin cu trn i tng Penicillium notatum. Nhng thi i khng sinh mi thc s bt u t nhng nm 1941, 12 nm sau pht minh ca Fleming khi penicilline c tung ra th trng, loi thuc khng sinh ny c tiu th rt mnh m. T my trm triu n v sn xut nhng u tin n nm 1949 th s lng n v a ra th trng hng thng ln 800 t n v. n nm 1944 Schatz, Bugio, Waksman tm ra streptomycine. n nm 1972, 4000 n 5000 loi thuc khng sinh c pht hin
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trong c cha 50 loi khng sinh c vn dng trong lm sng iu tr di dng n c hay kt hp, to nn mt lng khng sinh a dng c th chng cng lc nhiu loi bnh. Nh cng ngh sinh hc lng khng sinh vi s lng ln c to ra nh nui cy gene hoc cy nhng gene to khng sinh. Trc y ch c nm v vi khun nn c th sn xut khng sinh vi s lng ln trong mt thi gian ngn. Thuc khng sinh hin nay th c rt nhiu, mt s thuc c tc dng thn k trong nhng nm u mi tm ra tuy nhin mt thi gian sau th gim hn tc dng hay ht tc dng. Mt s thuc khng sinh mi c tm ra v danh sch ca n th ngy mt di hn. Tuy nhin vic s dng khng sinh i hi phi c s hiu bit. S dng ng liu lng v c khoa hc s em li hiu qu cao. Ngc li th s khng em li li ch g m cn rt nguy him. Hin nay liu php khng sinh vn l mt trong nhng phng php iu tr c hiu qu nht nhng thi gian qua thy nhiu bin chng virus, xut hin nhiu i tng mi. Nhiu vn cn c t ra v cn c gii p. V d penicilline c dng ln u khng thy c d ng nhng th nghim mi y cho thy c gy tai bin. Trong qu trnh pht trin ca y hc th cc bnh thn kinh v cc bnh tm thn ngy cng c ch mc du c pht hin t lu. T l ngi mc bnh kh cao, li nhng di chng v cng nghim trng, khng nhng i vi ngi bnh m cn vi ton x hi. Bn cnh cc phng php iu tri hin i nh phu thut, hun luyn di chng v thch nghi, tm l liu php v x hi liu php... th phng php iu tr bng thuc vn ng vai tr quan trng. My nm gn y, vic nghin cu v sn xut cc loi thuc v thn kinh v nht l tinh thn c nhiu cng hin ln lao. Nhiu loi thuc mi c p dng vo cng tc iu tr c kt qu tt, a mt s ngi bnh v vi cuc sng bnh thng. Cng vi s pht trin mnh m ca cng ngh gene, cng nghip sn xut vaccine ngy cng c nhn rng v ph bin l mt nhn t quan trng i vi chin lc y t v sc khe cng ng trong tng lai.
7. Cng ngh gene

Ngy nay ngi ta cng i su vo tm hiu nhng vn sinh ha ca gene. Nh hiu bit su sc v sinh hc phn t gene ngi ta c th bit rng con ngi c khong 35.000 gene v c ti 2% trng hp b
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bnh do khuyt tt gene. Ngy nay nh gii m c b gene ngi, lp ra c bn gene cho nn ngi ta c th gii quyt nhng trng hp b bnh do khuyt tt gene bng cch thay th cc gene b khuyt tt bi cc gene khe mnh. Khoa hc p dng cng ngh gene v cng ngh sinh hc bt ng vt, vi khun, nm men, virus sn xut nhng thuc cha bnh qu vi mt liu lng ln iu tr nhng bnh tt him ngho m trc y ngi ta ch to ra c vi mt lng qu t i. Bng cng ngh gene ngi ta to ra c chui t bo tit ra dopamine cha bnh Parkinson. Cng bng cch ny ngi ta xy dng phng php cha tr cn bnh Alzheimer (bnh l ln ngi gi). Ngi ta cy t bo thn kinh ln bao t vo no b ca Maribeth Cook, mt ngi ph n 34 tui l nn nhn tai bin mch mu no t 5 nm trc lm lit na ngi sau phu thut. Cook c th i li gn nh bnh thng, ni nng lu lot, phong v tm l bnh thng.. Jimmfimn 20 nm kh v bnh lit rung, khng th i li, ni nng v hot ng bng hai tay. Su thng sau khi ca phu thut cy t bo thn kinh Ln vo no b, Jimmfimn bt u i li c. Maribeth Cook v Jimmfimn tham gia t u ca chng trnh nghin cu lm sng v k thut iu tr chn thng no bng t bo thn kinh Ln. K l hn, chng trai 20 tui Robert Pennington b bi gan trm trng, sinh mng ca Robert ch c th c cu sng trong trng hp c ghp bung gan ln duy tr s sng v sut 3 ngy chng trai c cu sng nh bung gan ly t 1 gene ln. Vic cy ghp t bo lch sn xut insulin c th cu sng bnh nhn b bnh tiu ung do c th mt kh nng tng hp insulin. Ln nhn bn cung cp ni tng cho ngi- hin tng trong tm tay. Mt con ln nh mu trng, tai to tn l Gordy ra i ti phng th nghim ca trng i hc Tng hp bang Mitsuri (M), l s kin khoa hc ln gy chn ng khng km g s kin con cu Dolly. y l mt loi ln tai to c to ra t cng ngh nhn bn m ra mt trin vng ln cung cp ni tng cho ngi bnh c yu cu thay th mt s b phn trong c th. T trc n nay vic ghp ni tng ln cho ngi cha thnh cng v b phn ghp thng b c th nhn o thi bi ni tng cho khng ph hp vi gene ca c th nhn. Ln Gordy nhn bn thiu 2 gene khc phc c kh khn ny. 2 gene ny khng hot ng v b loi b ra. C hai gene ny th chng c nhim v sn sinh ra mt loi
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ng trong c th nh enzyme -1,3-galactoziltransfease, vit tt l ggta1. Cht ny l th phm gy ra phn ng o thi v vy khi loi b 2 gene ny ng thi loi b lun ggta1 do khng th khng cn c tc dng v s khng xy ra phn ng o thi b phn l v vy ni tng ca ln Gordy s khng b o thi. Mi y, mt ph n Canada b bnh cholesterol mu cao gp 10 ln mc nguy him thng thng v thng xuyn c cn au tim lng ngi do tc nghn mch vnh. Nguyn nhn l do thiu gene recepter ca LDL (low density lipoprotein) nn khng lm nhim v chuyn vn c choleterol v gan gy m huyt lm tc nghn mch vnh. Nh cng ngh gene a gene vo t bo gan, sc khe ngi ph n ngy cng mt ci thin hn. M vn l ni hot ng nghin cu gene din ra hi h nht hin nay. Cc nh nghin cu trng i hc Michijan ang th nghim thuc tim gene chng ung th v v ung th t cung ph n. Ti vin Nghin cu Ung th Quc gia M ngi ta th loi thuc tim gene chng ung th no. Gene ung th trong cng ngh sinh hc. C mt s dng ung th pht hin l do gene ung th. Phn ln cc gene ung th u l nhng alen t bin ca cc gene bnh thng trong t bo, nhng gene ny m ha v tng hp nn cc hormone. Mt s trng hp ch ring l 1 gene ung th gy ra ung th. Ngi ta thc hin thnh cng vic truyn loi gene ung th ny cho chut lm cho chut b ung th (1989) Mt s trng hp khc, gene ung th ch c th thc t gy nn ung th nu c gn vi mt s t bin khc hoc gn vi mt s nhn t trong mi trng. V cc yu t gn vi gene ung th nu trn c th c nghin cu, xc nh qua nui cy t bo, do ngi ta s dng cc ng vt truyn gene pht hin cc nhn t dn ti pht trin ung th hoc cc bnh l khc ngi. Nhiu nghin cu trn ng vt v c trn ngi, u xc nhn bn thn ung th v u c tnh l khng di truyn. Ngi ta ch pht hin c cc t cht di truyn (v kiu gene) i vi vic to ung th. Nm 1982, ngi ta tch c t bo ung th ca ngi, cc on DNA c th lm bin i cc nguyn bo si ca chut pht trin trong nui cy, t ang bnh thng n t bo ung th. Cc gene ny gi l gene ung th.
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Trc y, mt giai on kh di, ngi ta vn cho nguyn nhn ung th l do t bin gene, t bin NST (t gy, tng s lng v t chc...) Ngy nay, ngi ta c kinh nghim chng minh ung th khng phi c nguyn nhn t t bin gene, t bin NST m chnh y l cc hu qu ca ung th. Khi mt t bo bnh thng bin thnh t bo ung th, ung th c tnh th trong t bo y xy ra nhiu bin i su sc v ha, l. T bo ny khng cn chu s iu khin c t chc ca c th na. Trong t bo ung th xy ra t bin gene, t gy, tng gim s lng NST khng theo qui lut no c, c bit lm cho t bo c kh nng tng sinh sn, phn bo lin tc c trong iu kin nui cy cng nh trong c th ngi b ung th, ci m ta thng gi l di cn ca ung th. Trong thc nghim nui cy trong ng nghim ngi ta xc nhn virus c th bin i t bo lnh thnh t bo c tnh ngi. Vi khun than ang l ni kinh hong cho ton nhn loi th nay c cc nh khoa hc M xem l bo bi, cc nhn vin nghin cu pht hin cc protein bnh than sau khi bin i gene c kh nng tiu dit t bo ung th v khng lm tn thng n cc t bo khc. Trong c th ngi ta pht hin c im quan trng ny cho thy nhn loi c rt nhiu kh nng tm ra loi v kh tn cng hu hiu chng ung th. c tin hnh th nghim trn 100 con chut khi cho chng nhim 3 chng ung th khc nhau: ung th tht t chc mm, ung th da v ung th phi. Sau ly protein bnh than (thnh phn chnh ca vi khun bnh than) bin i gene chia lm hai ln tim. Kt qu t c gy sng st cho nhiu ngi, bi v cc t bo ung th gn nh b tiu dit ht v trong sut qu trnh th nghim th cc t bo lnh trong c th chut khng h b tn thng. Dng sng bn gene cha ung th. Mt nhm cc nh nghin cu M va tuyn b pht minh ra loi "sng di truyn gene" c th bn cc vin n cha DNA vo trong t bo chng cc khi u ung th. Bc s Aruna, T trng t nghin cu i hc Winsconsin (M) ni:"dng sng bn gene c u vit hn cc phng php a gene vo c th cha bnh ch n khng c hi, d dng xuyn qua vch t bo". Tuy nhin ng cng tha nhn dng sng bn gene ch thch hp cho vic cha tr cc khi u gn b mt da. Nhng nu ct b mt da th cng c th dng gene cha tr cc khi u nm su bn trong. Nm 2002, gio s Daniel thuc i hc Stanford (M) tin hnh th nghim dng virus cha tr cho 35 bnh nhn ung th gan
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giai on cui. Sau khi tin hnh cy virus vo gan th ngi bnh xut hin nhng biu hin ca bnh cm nh au hng, s mi v st cao. Theo cc nh khoa hc th y l biu hin ca cc phn ng ph ca liu php virus: "Cc phn ng ny nh hn nhiu so vi triu chng: bun nn, suy sp th cht v b rng tc ". Cc bnh nhn ny theo d on ca bc s ch c th sng ti a 6 thng. Song sau khi cy virus vo gan h sng c trn 2 nm. Qua siu m, cc khi u trong gan ca 35 bnh nhn ny nh dn hoc khng pht trin thm. Cc nh khoa hc ti i hc Southwestern medical center bang Texas (M ) cng tin hnh th nghim dng loi Adenovirus trong iu tr ung th bung trng. Nhm cc nh khoa hc ca May Oclinic tin hnh th nghim thnh cng trnh vic dng vaccine chng bnh si, c cha virus si iu tr cc ca ung th mu. Hin nay cc nh khoa hc ti Oclinic ny ang chun b cc bc th nghim trn ngi. Cc loi virus c s dung trong liu php gene c coi l nhng dn xut ca DNA. y l c s cc nh khoa hc dng virus bin i gene. Trong iu tr bnh ung th phi cc nh khoa hc pht hin ra rng trong DNA ca ngi mc bnh ung th phi c s thiu ht mt phn gene. H ly phn gene thiu ht ny cy vo virus c bin i gene. Qua ng th cc virus ny c ngi bnh ht vo phi, sau khi vo phi cc virus ny sn xut ra protein chng li ung th. Gn y, gio s David Sandersen cng vi cc cng s i hc Purdue thnh cng trong vic bin i gene virus Ebola, vn d l loi virus ny khng nhng khng cn nguy him i vi con ngi m cn tr thnh loi tn dc iu tr cc bnh v phi rt c hiu qu. Trng hp insulin tng hp t nhin trong c th khng , glucose phi thi ra ngoi theo ng nc tiu gy nn bnh i tho ng. Ngi b bnh i tho ng ch c cu sng nu b sung vo c th h mt lng insulin nht nh. Trc kia insulin t v t kh p ng c nhu cu ca nhng ngi b bnh. Ngy nay vi cng ngh sinh hc hin i ngi ta sn xut ra c nhiu insulin v r nn p ng c nhu cu cho nhng ngi b bnh i tho ng. Cng ngh sinh hc hin i sn xut ra insulin trong cc nh my t hon l vi khun E. coli. Trong dy chuyn sn xut ny, khu u tin l tch gene ph trch tng hp insulin trong c th sng c th t ngi hoc t Chut. Gene insulin cng c th c tng hp nhn to trong ng nghim. Sau l tch plasmid t mt loi vi
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khun, tc l vo t bo vi khun nhn (E. coli) . Khu tip theo l to dng DNA mang gene insulin bng cch cho gn gene ny vo plasmid to thnh mt DNA plasmid ti t hp mang gene insulin. y c s dng vai tr ca enzyme ct gii hn. Ngi ta chn mt enzyme ct thch hp i vi c DNA mang gene insulin v c plasmid. Enzyme ct si n DNA ni trn thnh cc on, ct m vng plasmid. Di tc ng ca mt loi enzyme ni, on DNA c mang gene insulin c chn ra ghp vo plasmid, sau plasmid c ng vng thnh mt plasmid "lai" c mang gene insulin. DNA plasmid ti t hp ny c chuyn vo nh my sn xut insulin ( E.coli). Bng cc phng php khc nhau nh dng CaCl2 tc ng lm mng t bo vi khun E. coli c th thm qua loi plasmid ni trn. Trong nh my ny DNA plasmid ti t hp c nhn bn gene, insulin trong cng c nhn bn v c truyn lin tip cho cc th h t bo vi khun qua c ch phn bo cc nhanh ca t bo vi khun E. coli, c sao m ri gii m tng hp ra hng lot insulin. Tt c u do b my di truyn ca t bo vi khun nhn iu khin. Cng ngh sinh hc vi gene khuyt tt . Bnh cm ic c nguyn nhn do vng no c lin quan n ting ni b tn thng. cha bnh ny trc y ngi ta phi ly t bo thai ngi (t cc trng hp ph thai) cy vo c th bnh nhn. Tuy c cc bin php lm thch nghi t bo c cy c th bnh nhn nhng nhiu trng hp vn gp kh khn do phn ng thi loi. Mt phng hng gii quyt bng CNSH hin i do cc nh khoa hc Nga vin Sinh hc gene ang theo ui v c rt nhiu kt qu l tch c cc gene c tc ng lm tng trng cc m thn kinh trn no b. Mt pht minh gn y nht c cng b l dng t bo thn kinh rui ci cy vo no bnh nhn cm ic bm sinh, bnh nhn teo no, nhn no, bi lit do no thiu mt loi t bo nht nh. Qu trnh th nghim c tin hnh u tin trn ch v sau l trn chut u thu c kt qu tt, chng minh r l cc t bo thn kinh rui ci c tc dng sinh ra cc cht kch thch sinh hc. Bnh nhn cm ic bm sinh c cy t bo thn kinh rui ci phc hi c kh nng ni, giao tip bnh thng. Tm ra loi gene khuyt tt gy nn cn bnh ri lon ngn ng tr em: Cc chuyn gia trung tm Nghin cu gene ngi thuc i hc Oxford (Anh) va qua cng b mt nghin cu mi trn Tp ch Nature
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s ra thng 10- 2001. Theo ngi ta pht hin thy NST s 7 trong tng s 23 NST ca c th ngi l nhn t quyt nh kh nng hc ngn ng ca con ngi. Trong loi gene ny c mt khuyt tt di truyn gi l FOX P2 gy cn s pht trin no ca bo thai lm cho tr sinh ra mc tt ni lp. Qua nghin cu gip cho khoa hc pht hin c c cu pht trin kh nng pht ngn ng ca tr th cng nh qu trnh tin ha v ngn ng nhng quan trng hn l hiu c c ch gy bnh t tm ra phng php khc phc chng ni lp. Nm 1998 cc nh khoa hc cng bt tay vo nghin cu nhng khuyt tt ca NST s 7 v pht hin c c nhng ngi khng c tin s gia nh mc bnh ri lon pht ngn ng nhng bn thn li c gene FOX P2 ny. Trong gene FOX P2 ny c cha 1 ha cht dng bc thang gi l guanine, cht ny c thay th bng adenine v s thay i cc nh c th gy ra nhng him ho rt ln cho con ngi. FOX P 2 l 1 thnh vin trong t hp gene to ra cc protein cho qu trnh sao chp gene v bin chng thnh th truyn tin RNA. y l mt trong nhng qu trnh phc tp, trong to ra cc protein cn thit duy tr cho c th con ngi tn ti v pht trin. Cc hp cht guanine do adenine thay th s lm gim s lng ca cc vt cht to nn th truyn tin RNA v cui cng nh hng trc tip n qu trnh pht trin no ca bo thai lm cho tr em b mc chng ngng, ni lp v nh trc tip n vic giao tip hay cn gi l cn bnh ri lon ngn ng c trng ( PLD). Nghin cu trn ca cc chuyn gia WCHC m ra mt trin vng mi trong vic tm ra nguyn nhn gy bnh c lin quan n di truyn v t tm ra nhng bin php ngn nga t b m sang con ci. Gn y cng b thnh tu ca cc nh khoa hc Php thuc trung tm Nghin cu Khoa hc Quc gia (CNRS) v vin Nghin cu Di truyn Phn t Paris pht hin nguyn nhn ca bnh bo ph do s xut hin ca gene t bin bta 3, l gene cm nhn adrenaline bng cc k thut cng ngh sinh hc hin i, ngi ta tm cch tc ng, khi phc hot ng ca gene do s khim khuyt v di truyn gii quyt "tn gc" chng bo ph trn c s phc hi li trng thi cn bng c th. Tht l mt dch v gene v cng ngh gene hp dn v si ng ang din bin t cc bnh c cong v ng nghim i n kt st v nh bng. Vi ny ca cng ngh gene, ngi ta rt lc quan s to c nhng nhn ti nh kiu Moza, Einstein..v to ra nhng con ngi p:" trong nh ngc trng nh ng".
8. Bin php th thai in-vitro
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Cch y 26 nm, (nm 1978), c th gii kinh ngc n cho Louise Brown, ngi u tin ra i bng phng php th tinh trong ng nghim. Hin nay trn th gii c khong 500.000 ngi ang sng cng ra i trong ng nghim. ng 20 nm sau, khi a tr u tin ra i trn th gii bng phng php th tinh trong ng nghim, ngy 30 thng 04 nm 1998, ba em b th tinh trong ng nghim u tin ca Vit Nam ct ting khc cho i. Tnh n nay, ti bnh vin Ph sn T D thnh ph H Ch Minh c 2000 b nh th t chng trnh ny (em b th 2000 ra i ngy 11-12-2005). l mt hnh trnh k diu, gian nan v xc ng m c nhng chuyn ngm li vn thy bi hi lm c vic th tinh trong ng nghim, mt iu qu mi m Vit Nam v mi mt: k thut khoa hc, d lun x hi, lut php, Bc s anh hng lao ng Nguyn Th Ngc Phng Gim c bnh vin Ph sn T D phi chin u vi mt s chu ng v kin tr gh gm cho c t ph mang tnh bc ngot trong lch s Y hc Vit Nam ny. Vic c mt a con trong gia nh l ni khao kht lm m ca rt nhiu ph n him mun, ni bt hnh, thm ch dn n tan v gia nh ca nhng cp v chng v sinh. Theo ti liu thng k ca Trung tm Brown Hall (Anh) ni a b th tinh trong ng nghim u tin ra i, cng b trong nm 1999 th t l thnh cng ca phng php th tinh trong ng nghim thng thng (IVF) l 30%, trong khi ti bnh vin T D t ti 38-40%. Vi nhng k thut th tinh trong ng nghim phc tp hn nh th tinh trong ng nghim bng k thut ICSI (bm tinh trng vo trng) ti B, ni u tin p dng k thut ny, hin t l thnh cng l 35,9% th ti bnh vin T D c thc hin trong 171 ca, m trn con vung 31 ca, t l thnh cng 41,5%. Th tinh trong ng nghim bng xin trng ca ngi khc th Brown Hall t 33% th ti bnh vin T D tht ng kinh ngc, t l thnh cng n 51% vi 111 ca thc hin th 56 ca thnh cng. Th tinh trong ng nghim ang hng v pha trc nhng cn rt nhiu vn cn gii quyt trc nhu cu v mn t cung, cho nhn tinh trng hin ang ng bng ch quyt nh ca php lut. y cha ni n vic th tinh trong ng nghim khng phi lc no cng sun s nh chng ta mong mun. Trong phng s khoa hc t Tp ch Th gii mi nm 2000 cng c bi vit Th tinh trong ng nghim; hnh phc i cng ni lo c vit: th tinh trong ng nghim dn n a thai v sinh non. Khi tr cho i khng thng ngy thng mang mt s bnh nh suy h hp, xut huyt mng no, thm ch pht trin tm thn v c bit l bnh vng mc mt gi tt l ROP
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(retinopathy of prematurity) hoc b RLF (retrolental fibrosis) dn n m hon ton. Th tinh trong ng nghim l th. Tuy nhin, i vi nhn bn ngi, c l mt trong nhng vn gy tranh ci nht l sao cho gi c bn sc ring ca tng c nhn c sinh ra trn i ny. Ngy 31 thng 7 nm 2001, Quc hi M thng qua o lut cm mi hnh thc nhn bn phi thai ngi. Mt s nh khoa hc lp tc ln ting phn i o lut ny. Xt v kha cnh y hc th vic nhn bn t bo gc trong phi thai s m ra nhng trin vng cha tng thy iu tr mt s bnh c gi l v phng cu cha. Nhng nu dng cng ngh nhn bn to nn mt con ngi hon chnh th li l iu hon ton khc. D th no i chng na, chng ta phi tha nhn rng, kh c th ngn c bc i ca cc nh nhn bn hc. Cn v tng lai ca cng trnh c mt khng hai ny th nh Gregor Pence, mt nh trit hc ngi M nhn xt: Nu mt a tr u tin ra i bng nhn bn khe mnh th chng ta s du ngay i v quen dn vi n. Nhng n li m yu, qut quo hoc thm ch cht th vic nhn bn s b cm trong vng 100 nm. Theo tin tc t Seoul, Hn Quc (12/2/2004), cc nh khoa hc Hn Quc nghin cu t 245 trng nhn thnh 30 phi. Phi pht trin t t bo gc v c kh nng phn chia thnh t bo cc c quan. Cc nh khoa hc Hn Quc khng c nh nhn bn v tnh hon chnh mt con ngi m trong qu trnh phn chia phi thnh cc c quan, cc nh khoa hc dng n trong vic cha tr mt s bnh him ngho. Gn y cc nh khoa hc Australia v M cng ang nghin cu t bo gc thay th cc c quan m con ngi b thng tn cc c quan .
1. 2. 3. 4. 5.
TI LIU THAM KHO i Duy Ban, L Th Cm Vn, 1994. Cng ngh gene v cng ngh sinh hc ng dng trong y dc hc hin i Nxb Y hc H Ni. Phan C Nhn- Trn nh Min,1997.Tm hiu cng ngh sinh hc hin i.Nxb Gio dc H Ni. Trng Vn Lung, 1995. Chuyn cng ngh sinh hc.T sch i hc Khoa hc Hu Nguyn Vn Uyn, Nguyn Tin Thng, 1996. Nhng kin thc c bn v cng ngh sinh hc, Nxb Gio dc H Ni. Bezborodov A.M., Moxolov V.V., Rabinovitch M.I., Nguyn Vn Uyn, Ng K Sng v nnk, 1994. Cng ngh sinh hc v mt s ng dng ti Vit Nam, Tp I, II. Nxb Nng nghip H Ni.
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6. Tp ch Th gii mi s 565, trang 3, trang 67.Tp ch Th gii mi s 566, trang 3, 6. 7. Tp ch Th gii mi s 571, trang 57. Tp ch Th gii mi s 565, trang 3, trang 67. Tp ch Th gii mi s 595, tr: 50, 56, 60. Tp ch Th gii mi 613, trang 3-7. Nxb Gio dc H Ni. 8. Tp ch Tri thc tr s 78, trang 87,88. Tp ch Tri thc tr s 80, Tp ch Tri thc tr s 93, trang 101. Tp ch Tri thc tr s 98, trang 71. 9. Bnh vin Bch Mai, 1968. Thuc iu tr cc bnh thn kinh. Nxb Y hc H Ni 10. Bo co khoa hc Hi ngh Cng ngh Sinh hc ton quc ln th nht thng 12/1999. 11. Bo co khoa hc Hi ngh Cng ngh Sinh hc ton quc 2003, H Ni 16-/12/2003 12. Bo co khoa hc Hi ngh Nhng vn nghin cu c bn trong khoa hc s sng nh hng y dc hc, Hc vin Qun y ngy 28 thng 10 nm 2004. 13. Albert Sasson,1988. Biotechnologies and development Cng ngh sinh hc v pht trin. Ngi dch: Nguyn Hu Thc, Nguyn Ln Dng v mt s dch gi khc. Nxb Khoa hc & K thut H Ni.
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CNSH vi bo v sc khe cng ng Truong Vn Lung
Chng IV
S kt hp cng ngh sinh hc c truyn v cng ngh mi trong vic nng cao sc khe con ngi
y l vn cn ang mi m v ang c nhiu nc s dng nng cao kh nng cha tr bnh cho con ngi. y cng l mt hng pht trin mi cho ngnh y hc th gii v c trin vng ln.
1. Kt hp chn on ng y-Ty y nh hng bnh

Mt trong cc phng php hin i ha y hc c truyn l dng phng php, k thut ca Ty y chn on, t thy thuc quyt nh nn cha theo Ty y hay ng y. Nhiu bnh cn phi theo Ty y mi c kh nng cu cha hoc cu sng nh cc bnh cu khn cp, hn m, bnh cn phu thut, bnh nhim trng nng. Nhiu bnh mang tnh xc nh r nguyn nhn hoc c tn thng r rng, cha theo Ty y c kt qu r rt nh lao phi, lot d dy, t trng, vim phi, ung th pht hin sm, Cc phng php hin i nh ni soi d dy, i trng siu m, siu m qua ni soi, CT scaner, chp ng mch vnh, thng tim, ct t, chn on sm trong khi ng y khng c kh nng xc nh. Ngc li, ng y, theo T chc Y t Th gii, c kh nng cha tt cc bnh mn tnh v tiu ha, thn kinh, au nhc, cc bnh chc nng, (khng c tn thng r rng). Thuc b ng y, thuc kch thch n ung c tc dng tt. Chm cu, bm huyt c tc dng tt cho bnh au dy thn kinh ta, lit dy thn kinh s 7, gy t. Mt s bnh cha theo ng y r tin, hiu qu cao nh vim d dy, xung huyt, bnh i trng chc nng. Ngy nay nhiu thy thuc ng y tin hnh khm bnh theo phng php lm sng, xt nghim, siu m song song vi chn on ng y theo th bnh. V vy, cc lng y thu c kt qu tt trong iu tr.
2. Kt hp ng yTy y cha bnh

Nh phng php phn tch hot cht chnh xc, th nghim trn sc vt m ngi ta pht hin ra tc dng mi khc hn trc y. Th d: t trch, h th c cha chng m trong mu cao, kim tin tho cha thn, du ht tiu cha hen, bch hoa x thit tho cha ung th. Khi pht hin thuc c tc dng tt, khng c th kt hp cng thuc ng-Ty y cng ngy hoc cng lm thnh mt vin thuc u c tc dng tng ln. V d: thuc tiu kht hon gm c thuc ng y v
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mt loi thuc Ty y nh Daonil. Thuc tim phong hon gm thuc ng y v mt cht thuc Ty y cha gim au, chng vim khp. Sai lm ln nht khi kt hp ng-Ty y l quan nim sai v ph tng, c quan. Sai lm th 2 l khng bit r tc dng ca thuc nn p dng sai. Mt s ngi quan nim thuc Bc, thuc Nam khng c, gy mt, nn dng di ngy hay liu cao. S ngi ng c thuc ng y vn thng xy ra. Mc nng th gy hn m, tt huyt p, suy gan, suy thn, Mc nh hn, gy vim gan do ng c, nn ma, suy thn mn tnh din bin chm. Nguyn nhn gy ng c thng do truyn ming, t dng khng bit r tc dng hoc chn on sai, hiu sai chn on ng-Ty y. C ngi b hn m do ung v cy v sa hi vng cha tiu ng. Nhiu ngi b ng c mt c trm khi ung (thc ra ch dng ngoi nh bi da). Khi kt hp ng-Ty y trong mt t, mt ngy, thng thng theo nguyn tc khng c, khng lm mt tc dng ca thuc kia, khng gy bin chng nng hoc tc dng ph. Phi bit chc, hiu r tc dng ca tng loi thuc kt hp. V d: vitamin c th kt hp vi nhn sm, c tc dng b dng. Ngi ta pht hin ra interferon dng cng vi tiu si h thang gy bnh gan nng hn. Ng v t dng chung vi sorbetol gy tiu chy. Thuc tin kht hon c thm thuc Ty h ng huyt li dng thm thuc h ng huyt khc nh diabinese s gy tt ng huyt. Khng kt hp hai loi thuc c vi nhau. V d: l kh kt hp vi thuc c Ty y, hai thuc lm tng tc dng ca nhau khi dng liu cao v ngay c khi dng ng liu, gy bnh nng hn. Hoc laxix dng chung vi ru ng, m , r c tranh gy ra tiu nhiu, mt nc, ri lon in gii. Khng dng qu nhiu thuc khi khng bit r tc dng, mt thang thuc ng y thng thng c t 5 n 10 v, trong khi cha bit r ht tng v li dng thm thuc Ty y c th gy ng c cp hoc ng c t t m khng nhn ra. Cc bnh ng c thng gp nht l suy gan, suy thn, chy mu nng do dng thuc tim vo bi tr. Phi bit r chn on ca Ty y hoc ng y kt hp thuc. Nu khng bit bnh lot i trng m li dng i Hon s gy tiu chy nng hn. Tm li, khi kt hp hai loi thuc ng y v Ty y nn kt hp vic lm tng tc dng cha bnh nhng khng gy tc dng ph. Cn phi gim liu, phi theo di khi dng chung, khng dng nhiu thuc khi cha r tc dng. Khng dng cc loi d gy c vi nhau. Cn da vo
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hai chn on ng v Ty y la chn thuc. Khi phi hp thuc nn cch mt thi gian theo di. Phng php hin i ha y hc c truyn ang c tin hnh nh dng khoa hc hin i phn tch, chn on, phn bit bnh t chn ra mt phng php iu tr thch hp. Chn thuc c tr t dc tho, tm ra tc dng mi, tm ra hot cht chnh ang c p dng rng ri. S kt hp gia hai chn on ng-Ty y trn mt ngi bnh s trnh c sai lm. Mt iu trong nhiu bnh vin hin nay hay s dng l trnh hi sc khe cho con ngi khi gy m trong nhng ca m, ngi ta dng phng php chm cu. Ch cn mt s kim chm vo cc huyt th ngi bnh nm trn bn m thy ht, bit ht nhng thao tc ca bc s ang m cho mnh nhng khng h thy au n. ng nhin, mt s ngi yu bng va th h rt s trc nhng thao tc ca bc s khi a con dao vo rch da tht mnh. Chng ta khng nn qun li dy ca bc H: Thy thuc Ty y phi hc ng y, thy thuc ta cng phi hc Ty y th mi tt. Mt iu cng cn phi quan tm trong giai on hin nay l iu
tra hot cht sinh hc (bioprospecting).

Theo Ian Paterson (Gio s i hc Cambridge, M) ni: Th gii cc c th sng tri qua mt thi k di u tranh sinh tn pht trin c cc kh nng v cng phc tp v c th ng dng trong rt nhiu kha cnh ca y hc. B gene ca cc sn phm t nhin khng bao gi ph thi gian m khng to ra cc hp cht mi khng tc ng ti cc h thng sinh hc. iu tra thm d hot cht sinh hc trong sinh vt sng c th mang li li ch to ln v y dc hc v thng mi. ng nhin y l lnh vc u t mo him, c th tht bi v cng c th thu li nhun rt ln. V th, cc quc gia ang pht trin thng khng tim lc v ti chnh, v trang thit b v c bit v chuyn gia trin khai cng vic ny, mc d ti nguyn thin nhin v tnh a dng sinh hc ca cc nc ny ht sc phong ph v to ln. Hot ng iu tra thm d hot cht i hi s kt hp cht ch gia kinh nghim c truyn trong vic s dng cc ng vt, thc vt, nm v vi sinh vt lm thuc cng nh nhng trang thit b hin i ca phn tch ha hc v sinh hc. Cng vic thng bt u vi nhng nhm cy con c y hc c truyn s dng, sau m rng mt cch c h thng n cc khu vc vn Quc gia, tip n l cc h vi sinh vt sng trong cc h sinh thi c bit nh rng ngp mn, k sinh trn cy dc liu, trn rong, to bin, V phng php tin hnh trc tin l iu tra tm hiu kinh
112
nghim s dng thuc ca ngi dn, sau dng cc phng php hin i tch chit, thng l cc loi dch chit, c a v phng th nghim tin hnh sng lc qua cc php th c trng cho cc nhm thuc nh khng khun, khng virus, cha st rt, cha ri lon thn kinh, cha bnh lao, c ch sinh trng ca t bo ung th. Rt c th phi phi hp nhiu phng th nghim t chc sng lc mt cch c h thng v c hiu qu. Nhng mu dng tnh s c phn tch ha hc nh sc k cao p, in di, khi ph cui cng i n hot cht. V d, gn y ngi ta tm thy cht chng HIV gi l conocurvone trong r cy Cotinus coggygria. Rt nhiu quc gia pht trin hoc cng nghip ha (NICs) v cc cng ti ha dc cht mun u t cho loi cng vic ny. Ti vin Nghin cu Eisai (bang Massachussets, M) cc nh khoa hc iu ch c cht k hiu Halichondrin B, dn xut t mt loi bt bin sng di tng nc su trong vng bin New Zealand, c cho l c bit cng hiu trong vic iu tr ung th giai on cui. Cng ti Bristol-Myers Squibb ang th nghim mt loi thuc mang tn Ixabepilone , m phng theo mt loi vi khun c nhiu trong t mn bnh thng nhng c th cha khi bnh ung th. Tp on Wyeth gn y gii thiu ra th trng sn phm mi bit dc Rapamune phn lp t mt loi giun t ca o Easter c tc dng phng nga hin tng thi loi trong ghp thn. Rapamune cng l ch phm chy nht ca hng trong thi gian qua. (theo Tp ch Th gii mi s 664, thng 12 nm 2005 trang55-58). Th gii sinh vt tht a dng v phong ph. Nu khai thc c th gii ny d ch mt phn nh, y hc cng s thu c nhng thnh cng vt bc phc v c lc cho sc khe cng ng. V th, nhiu t chc quc t cho ra i nhng vn bn nu nguyn tc chung v bioprospecting, trong c vic tha hng nhng kt qu s dng ca phng php c truyn kt hp nhng phng php hin i to ra nhiu sn phm dc hc nhm cha tr ngy cng tt hn nhng bnh m trc y cho l v phng cu cha. TI LIU THAM KHO (Tham kho ti liu chng III )
CNSH ch bin cc sn phm sau thu hoch
113 Trng Vn Lung
Chng V
Cng ngh sinh hc trong vic to sn phm hu c thc phm

1. Th gii sinh vt l ngun nguyn liu v tn cho cng nghip ha thc phm
T cui th k XIX, mt trong nhng ngi sng lp ra mn ha hc tng hp, nh bc hc ni ting ngi Php Marcelin Berthelot khng nh: Vn cc sn phm dinh dng l vn ha hc. Khi no thu nhn c nng lng mt cch r tin th thc hin c vic tng hp cc sn phm dinh dng t carbon (ly t kh carbonic) t hydrogen (ly t nc), t nitrogen v oxygen (ly t khng kh). Cc cng vic m cho n nay cy ci lm c nh nng lng mt tri. Chng ta trong tng lai khng xa s thc hin c. Bc tranh do Berthelot v ra, n ngy nay, vn l mt cnh in vin vin tng i vi chng ta. Dmitri Ivanovitch Mendeleiev sng sut cho rng, hin thc nht vn l sn xut cc nh my sn phm dinh dng nh sinh vt. Th gii sinh vt vn l ngun cung cp ha cht trong t nhin. Cc nh sinh vt hc v ha hc trong s hp tc sng to lm nn nhng qui trnh cng ngh hin i s dng c hiu qu nht. Th gii sinh vt dng lm nguyn liu cho vic ch to ra cc ha cht khc nhau phc v cho con ngi.
Ethanol Protein Keylit Sorbite Vitamin Thc n gia sc Cc dc phm Acid citric Fructose Xylose Agar Alginate Sn phm khc
Trng trt Lm nghip Cc sn phm quang hp
Du m
Sinh khi trn cn Sinh khi di nc
Kh t t nhin
Nng lng Ethylene Propylene Benzene Xylene Phenol Methanol H2 NH3 Acetylene Than cc Naphtalene Protein Furfurol Sn phm khc
Than
Hnh V.1. S cc sn phm ha hc c hnh thnh t thc vt
114 Trng Vn Lung
Tht vy, t nm 1950, cng nghip du m pht trin trong c ha du ch to cc ha cht t du m, m thc cht du m v than u t sinh khi t nhin do qu trnh quang hp to nn v ha thch theo thi gian vi lp di mt t. Ngoi ra, trong thc vt cn cho nhiu sn phm ha hc khc. C th tm tt theo s c trnh by sau: Trong cng nghip cung cp du bo: tinh du, hng liu, tannin lm mu nhum, rezin, du nhn, cao su, si, bt giy. Cc cy thuc cho dc phm, cy c cht c, thuc c, thuc tr su. Cy lm nha m: lm cht t, nhin liu. Mt s rong bin cho agar, alginate, carageenan. M c voi, mi thm c cung, x hng chn, mi vani, cht cho mi vi cht gi hng dng trong gii kht,v.v. Ngoi nhng cht t nhin c con ngi s dng t lu cn c nhng cht qu him m trong ha hc khng th ch to c.ngi ta phi nui trng trong iu kin cn thit cho qu trnh sng ca chng (thnh phn v t l vi lng thch hp, iu kin kh hu thch nghi). l nhng cy con c sn ca tng vng m ni ny c, ni khc khng c hoc nu c th cht lng sn phm thua km. V d: nhn sm Triu Tin c mt s vng trng c loi tt. nc ta c tam tht, trm hng, qu Thanh (Thanh Ha), qu Tr Mi (Qung Nam). Gn y ngi ta pht hin trn nh ni Ngc Linh (Kontum) c loi nhn sm (ngi ta t tn l nhn sm Ngc Linh), phm cht cn tt hn c nhn sm Triu Tin. Php c loi nm Truffe, gi c lc ln 4.000 Francs/kg. Nm Melanosporum tuberculum sng vng ni vi, sng cng sinh vi cy si (Quercus) mc t 1-2 dm. tm ging nm ny, ngi ta dn ln i nh hi. Nm ny to hng t nhin khi nu gan vt, ngng. Shikonin l cht c mu dng lm cht ch mu. Cht ny ly t r cy Lithospermum erythrorhizon c Nht Bn, Triu Tin. Cy ny trng 5-7 nm, chit r ly c 1-2% cht kh, gi 1 kg l 4.500 USD. Nhu cu tiu th shikonin cao. Nht Bn phi nhp thm ca Trung Quc, Triu Tin. Hin nay, Nht Bn dng k thut nui cy m v t bo nhn ging, chn dng t bo c t l shikonin cao (14-15%). Ngi ta chn dng t bo c mu m th t l shikonin cng cao. Ngi ta dng ni ln men 750 lt nui t bo trong 15 ngy thu c 5 kg. Nh phng php ny m khi lng thu c nhiu v chn dng c cht lng cao. Bng con ng nui cy m v t bo hiu qu gp 800 ln so vi nui trng t nhin. Nh CNSH can thip vo m nhiu sn phm qu c nhn ln nhanh, do vic thu nhn cc sn phm qu c nhiu.
115 Trng Vn Lung
Vic hiu bit cc c ch di truyn v k thut thay i hng sn xut v m ra nhiu trin vng tt p. Bungarie, hoa hng c to gene, khng ch gene to mi thm biu hin sm, thu c nhiu cht c mi hng khc nhau v r tin hn nhiu ln so vi vic thu hng t nhin. Ngoi ra, trong nui cy c th t ng v iu khin c nh in ton (to pH, m, ).
2. S chuyn ha sinh hc l c s khoa hc ca cng ngh ch bin to ra sn phm hu c thc phm

CNSH, cc sn phm ca s tng hp sinh hc v cc qu trnh ln men chuyn ha vt cht gn lin vi nhng iu mi m v c khi l bt ng khng nhng ch i vi nng nghip, lm nghip, y hc (phn mt v hai). Cng ngy cng thy r kh nng ng dng c th v rng ri chng trong cc ngnh ha hc v nng lng hc. Vic sn xut cc acid hu c, acid amin, con ngi ch to ra n s dng kh sm. Cha ng ta t xa cng bit mui da, lm gim, lm tng, chao s dng trong i sng hng ngy. Acid citric c sn xut t chanh.. Sau i chin th gii ln th nht (1920), ngi ta sn xut bng con ng vi sinh vt. Sau ngi ta cng sn xut nhiu acid amin khc nhau. T nm 1909, Nht Bn bt u sn xut acid amin l acid glutamic. 2.1. S chuyn ha sinh hc (biotransformation) v cc ng dng trong sn xut v cng nghip Cc sinh vt c kh nng thc hin nhiu loi phn ng ha hc khc nhau . Cha k n nhng phn ng ha hc ng vt v thc vt, trong vi sinh vt ngay t nm 1959, ngi ta pht hin thy hn 1.500 phn ng khc nhau. Nhng phn ng chung bao gm: - Phn ng oxyha: decarboxyl ha cc acid amin - Phn ng kh: phn ng 2NH2 + 3H2 2NH3 - Phn ng carboxyl ha to nhm COOH cc acid hu c - Phn ng mt amin (desamination) - Phn ng to glucoside xy ra khi tng hp tryptophan - Phn ng thy phn - Phn ng methyl ha (gn nhm CH3) - Phn ng ether ha, ester ha - Phn ng mt nc - Dismutation: khi tng hp cc cht ch c chuyn i v to ra cc hp cht trung gian, sau c bin i to thnh cht mi
116 Trng Vn Lung
- Phn ng kt hp (condensation: ngng t, c c) - Phn ng amin ha: - Phn ng acetyl ha to acetylCoA - Phn ng amylin ha: ct bt thnh ng Cc phn ng ny c thc hin trong cc ngnh cng nghip sn xut ra cc sn phm cn thit. Mt trong nhng v d in hnh l s dng cc sinh vt thc hin cc phn ng trong ch bin D-sorbitol thnh L-sorbic:
CHO H-C-OH OH-C-H Ha hc H-C-OH H-C-OH CH2OH Glucose CH2OH CH2OH H-C-OH Acetobacter C=O OH-C-H suboxydase OH-C-H H-C-OH H-C-OH H-C-OH OH-C-H CH2OH CH2OH D-Sorbitol L-Sorbose
Dng biotransformation L-sorbose L-acid ascorbic (vit.C) Nh vi khun chuyn phn ng D-sorbitol thnh L-sorbose m sn phm c h gi thnh so vi thc hin phn ng bng ha hc. Vic sn xut cortison cht tr thp khp, au nhc c tng hp ha hc bn ra th trng. Nu bng con ng ha hc th phi tri qua 37 cng on. V vy gi thnh cao (gi 200 USD/g). Nh s kt hp gia ha hc v vi sinh vt hc m qui trnh sn xut ny rt ngn cn 11 cng on v gi thnh ch cn 0,68 USD/g m khng cn n nhit v p sut cao. Rt nhiu cht khng sinh hin nay dng trong iu tr, ngi ta sn xut bng con ng ha hc kt hp vi sinh vt hc. 2.2. Sn xut acid hu c * Acid citric: Nhng nm 1920, ngi ta sn xut acid citric bng cch dng Aspergillus niger. Ngi ta cng tuyn chn nhng ging Aspergillus c nng sut cao sn xut c hiu qu. C cht cho qu trnh ln men ny l mt r ng, nui pH thp (pH=3). Di hot ng ca Aspergillus, pH chuyn sang dng acid (pH=1), sau ngi ta chit ra c acid citric. Trong qu trnh ln men, dng Aspergillus niger chuyn c 90% ng thnh acid citric. Ngi ta cho CaCO3 to thnh ta calcium citrate lng xung, sau cho tc dng ta vi H 2SO4 to thnh acid citric v CaSO4 lng xung. em acid citric c, kt tinh. Trong qu trnh ln men, b mt to thnh mt lp bo t en dy.
117 Trng Vn Lung
Sau ny ngi ta sn xut acid citric bng cch nui Aspergillus trn parafin du m. * Acid gluconic: c mt thi gian, ngi ta sn xut acid ny bng con ng ln men vi sinh vt t glucose di tc dng ca enzyme glucosoxydase: enzyme Glucose acid gluconic Glucosoxydase l mt enzyme cn khng kh, v vy trong sn xut cn sc kh mnh. Ngi ta ta acid gluconic bng CaCO3 to thnh calcium gluconate (dc phm dng cm cho tr em). Hin nay, ngi ta vn sn xut bng con ng ha hc l ch yu, v vi sinh vt rt d b nhim v chn chng tng i kh. Enzyme glucosoxydase s dng bo qun thc phm. Cho vo bia mt t enzyme ny s bo qun lu hn. Bi v, trong bia cn mt t ng glucose. Chng c ln men thnh acid gluconic s loi c O2 v bia hi chua mt t nn t b nhim v gi c lu hn. * Acid itaconic: c sn xut t Aspergillus itaconicus v Aspergillus terreus. C cht ca n l mt r ng, cht tinh bt, ng. Acid ny c dng trong sn xut polymer v c dng trong sn xut len nhn to. Khi cng nghip ha du pht trin con ng sn xut ny b ngng. * Acid lactic: c sn xut ch yu l dng vi sinh vt ln men lactic. Cng nghip ha du pht trin th sn xut acid lactic bng con ng vi sinh vt cng b ngng. 2.3. Sn xut acid amin * Acid L-glutamic: ln u tin vo nm 1909, hng Ajinomoto Nht Bn sn xut acid ny. Tht ra, bt ngt c sn xut t nm 1908 bng cch thy phn gluten ca bt m. Ngi ta dng Micrococcus glutamicum chnh xc hn l Brevibacterium glutamicum. Trc y sn xut c 40g/l. Hin nay nh chn lc dng c nng sut cao nn sn xut c 120g/l. * Lysine: cng c sn xut lng ln b sung vo thc phm. Chu trnh ca chng l: L-aspartic L-lysine L-aspartic Homoserine Methionine semialdehyde Threonine Isoleucine Trong sn xut cn c ch con ng v to thnh lysine c nhiu hn. Sau y l kt qu ca vic sn xut acid hu c v acid amin.
118 Trng Vn Lung
Sn phm Acid glutamic Acid citric Lysine Acid lactic
Gi n v (USD) 3.5-4,0 1,5 5,0-5,5 1,0-1,5
Tng sn lng (tn/nm) 300.000 300.000 40.000 40.000
2.4. S c nh enzyme v c nh t bo S dng cc enzyme: cc enzyme sn xut bng con ng vi sinh vt hoc tch t vi sinh vt c s dng kh rng ri hin nay. Trong thc t ngi ta pht hin ra 2.000 enzyme khc nhau; cho n nay, ngi ta thu c 200 loi enzyme v s dng rng ri trong sn xut S dng rng ri nht l cc loi amylase v proteinase. Sau y l mt s ngun v ng dng ca enzyme:
Tn enzyme -amylase -gluconase Glucoamylase Glucoisomerase Ngun Aspergillus oryzae Bacillus amyloliquefaciens Aspergillus niger Aspergillus niger Artinobacter sp. Bacillus sp. ng dng To siroglucose H tinh bt Dch ha trong ln men bia Thy phn bt Fructose cao cp Siro t bt ng c fructose cao cp
Vic ng dng enzyme vo cng nghip ngy nay cng c m rng. Gn y pht hin ra c ba loi enzyme x l sn phm du m to polymer, tnh ra trong thi gian ti li nhun c th ln ti 50 t USD. Sn phm ph v cht thi cha carbohydrate c th c chuyn ha bng cch ln men nh cc vi sinh vt thng thng hay bng cc qu trnh CNSH. V d: r ng to thnh t nc ci, sau khi kt tinh ng ma v c loi khi cng on ch bin khi nng ca ng tr nn qu thp. Ngoi ng ra, trong c sulphite, sodium carbonate v cc mui magnesium (c bit vi ng c ci). Tuy nhin, s ln men r ng khng chuyn ha c tt c ng st li. Hemicellulose gm 10% sinh khi g thng v 20% sinh khi g ca cc cy l rng, trn 30% rm r v li ng. Thy phn hemicellulose to ra xylen v xylose. Sulzer (Thy S) tch xylose t cht thi giu sulphite ca bt g c th thu 80 kg xylose/1 tn cht thi g hoc 120 kg xylose/1 tn b ma. Cho ln men trc tip hexose v pentose ly t dch thy phn cellulose v hemicellulose nh vi sinh vt phn hy cellulose to thnh ethanol th vn hp dn hn sn xut cn t tinh bt ng cc. S phn gii
119 Trng Vn Lung
v chuyn ha cellulose v hemicellulose bng vi sinh vt sn sinh ra ethanol v cc nguyn liu khc cho cng nghip ha cht (furfirol, phenol, cresol) c th ly ra c 20.000 tn ethanol v 22.000 tn furfirol t 200.000 tn rm c bin i hon ton. Nhng sn phm to thnh trn u do cc enzyme thc hin cc phn ng sinh ha trong c cht. Song c nhc im l chng d b mt hot tnh hoc gim hot tnh sau mt s ln s dng. Nu tch chit cc enzyme ra s dng th thng kh khn v tn km. V vy, hin nay ngi ta tin hnh c nh enzyme, ngha l gn enzyme vo mt chui no s dng nhiu ln. * Gn l hc (physical assosiation) kt hp c hc. S dng nhng vt xp (gch, snh s, gm), enzyme chui vo v gn vo . Thng ngi ta cho vo mt ct trn c mt s cht no cho phn ng. Phng php ny thc t t s dng v enzyme d gn vo th cng d tch ra, do , d b ra tri i. * Phng php ni nhng lin kt cng ha tr: mt u ca enzyme gn vo c cht, u kia ca enzyme vn t do.V vy chng vn c hat tnh bnh thng. V d: dng c cht l glutaraldehyde hay toluen diisocyanate thc hin phn ng bng nhng lin kt cng ha tr. Phng php ny t c s dng.
Glucose
Enzyme
glucosoisomerase
Fructosse * Phng php nht (entrapment): nhng polymer c dng si v nhng phn t enzyme ta ra nm nhng cht nn dng nht chng. Cc cht nn thng l cht xp, thng dng l collagen, gelatin, agar, alginate, carageenan, polyacrilamide, cellulose, triacetal, polystyrene. Ty
120 Trng Vn Lung
theo mc ch m s dng. u tin c th dng monomer m trn vi enzyme. Enzyme s nm vo trong. Sau dng alginate nu chy ra, cho c li, cng enzyme lm thnh vin. Cho vin vo cc ct ri cho dch alginate i qua. C nhiu loi enzyme c th dng s dng hng nm tri. Mc du to enzyme c nh tn km nhng s dng c nhiu ln. Hiu qu s dng enzyme cao. V d: Tinh bt
amylase amylase
Glucose
Glucosoisomerase
Fructose
Khi c k thut thut c nh, vic to fructose l phn ng thun nghch, nu fructose to thnh nhiu s phn ng tr li thnh glucose. S dng enzyme c nh glucosoisomerase ln ct dung dch glucose qua. Phn ng to thnh fructose chy ra ngoi ct. Nh vy m c th thu c dung dch fructose c n 90%. Enzyme c nh cn c th s dng c nhiu ln, trnh c c ch ngc v s dng c lu hn (nu nh khng s dng lin tc cng b nhim). Bng phng php ny, M hng nm thu c 4 triu tn. C nh t bo: trong nhiu trng hp, vic chit enzyme gp nhiu kh khn v tn km; nu dng c t bo th tn km hn. Vic c nh t bo cng ging nh vic c nh enzyme, c th dng cc phng php c hc, gn bng lin kt cng ha tr hoc nht. Vic c nh t bo cng c s dng t lu. Nh vic lm gim c truyn bng cch cho con gim bm trn mn ca, dch ng, ru, s to thnh gim. Nguyn tc l cho t bo bm vo mt vt no s dng n c nhiu ln trn vt c nh. V d: c nh t bo Saccharomyces serevisiae ln men. Ngi ta pha alginate lng trn ln t bo nm men cho thm CaCl2 alginate to thnh calcium alginate, cho qua li to vin. Khi dng cng nh i vi vic sn xut fructose trn, cho dch ng i qua ct c cha vin men. Qu trnh ln men ru c thc hin. Phng php c nh ny, t bo c nh c s dng nhiu ln. i vi t bo thc vt, ngi ta cng c nh bng phng php ny. Bi v, vic tch enzyme ra khi t bo thc vt th kh hn. Chng thng c thnh t bo dy bng v cellulose. Ngi ta s dng bng phng php c nh t bo to ra cht ta cn th c li hn. qu trnh c nh c kt qu, cn ti cc yu t sau y: - Enzyme phi n nh trong nhng iu kin din ra phn ng.
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- Nu c th c th cc cht tham gia phn ng to lin kt ngang s ch yu ch tng tc vi nhng nhm chc nng nm ngoi tm hot ng ca enzyme. - Nu iu kin trn khng thc hin c th cht tham gia phn ng to lin kt ngang phi c kch thc ln, khng cho php n xm nhp vo tm hot ng ca enzyme. - Tm hot ng phi lun lun c bo v (nu thc hin c) bng cc phng php khc nhau. Th d enzyme c cha nhm SH th cn phi x l s b n bng gluthatione hay cysteine v ch ti hot ha enzyme sau khi gn n vo cht mang. i khi c th che tm hot ng bng cch b sung vo hn hp phn ng c cht c bo ha bi enzyme. - Bin php ra tch enzyme khng cn gn, khng gy nh hng xu ti enzyme c gn. - Khi la chn h c nh, cn phi ti phn ng c th. Th d: tht v ngha nu bng con ng l hc ta a enzyme vo gel mang v s dng n xc tc phn ng phn r polymer cao phn t nh polysaccharide chng hn. Cng nh vy, cc polyanion s khng c vai tr ng k nu l cht mang enzyme xc tc bin i c cht anion thnh sn phm cation, c bit l, nu nh enzyme ny li cn b c ch bi sn phm ca phn ng. - Cn phi tnh n cc tnh cht c hc ca cht mang. c bit, iu ny lin quan ti nhng cht mang c s dng trong nhng ct ln.
1. 2.
3. 4.
5. 6.
TI LIU THAM KHO L Don Din, 1994. Cng ngh sau thu hoch trong nng nghip Vit Nam: Thc trng v trin vng. Nxb Nng nghip H Ni. Trng Vn Lung, 1995. Chuyn Cng ngh sinh hc. i Hc Hu Trng Vn Lung, 2002.Cng ngh sau thu hoch. (H hp vi vic ch bin v bo qun thc phm). T sch i hc Khoa hc Hu. Phm Vn Ty, 2001. Min dch hc. Nxb i Hc Quc Gia H Ni Nguyn Vn Uyn, Nguyn Tin Thng, 1996. Nhng kin thc c bn v cng ngh sinh hc, Nxb Gio dc H Ni. Trn Cm Vn, Bch Phng Lan, 1995. Cng ngh vi sinh v bo v mi trung. Nxb Khoa hc & K thut. Trung tm Giao lu quc t v Vn ha Gio dc v Khoa hc (CCES), H Ni.
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7.
Sasson Albert,1988. Biotechnologies and development Cng ngh sinh hc v pht trin. Ngi dch: Nguyn Hu Thc, Nguyn Ln Dng v mt s dch gi khc. Nxb Khoa hc & K thut H Ni. 8. Bezborodov A.M., Moxolov V.V., Rabinovitch M.I., Nguyn Vn Uyn, Ng K Sng v nnk, 1994. Cng ngh sinh hc v mt s ng dng ti Vit Nam, Tp II. Nxb Nng nghip H Ni.
CNSH Ch bin cc sn phm sau thu hoch
128 Truong Vn Lung
Chng VI
Cng ngh sinh hc ng dng trong khai khong v sn xut vt liu

CNSH ng dng trong khai khong v sn xut vt liu c nhiu c im tng t nh CNSH to sn phm hu c thc phm. Song, CNSH ng dng trong vt liu ngoi vic ch to, khai thc vt liu, cn l cng ngh dng thu hi cc cht v x l mi trng. C th sinh vt c th to ra nhiu loi vt liu qu m n nay con ngi cha th bng cch tng hp ha hc no thay th n.V d vic to thnh xng trong c th ng vt do thc n bin i m thnh; xng l vt liu kh thay th, ngi ta khng th dng kim loi th xng. Gn y ngi ta s dng vt liu thay th xng l san h bin. y l thnh cng bc u trong CNSH phc v sc khe cng ng m chng trc chng ti cha c dp cp n. ng v c im sinh hc m ni, cc o san h bin, t ong th cha c mt lu i no c th so snh c kin trc ca chng. Bi v, l nhng biopolymer, sn phm vt liu ca sinh hc.
1. S bin i, tch ly v c nh cc kim loi

Do nn nhim mi trng nn mt vn t ra l phi x l cht thi, lm sch mi trng, lm sch nc y l vn ang c ch gii quyt ca nhiu nc. C nhiu cch gii quyt: x l bng phng php vt l hc, ha hc; c th x l bng phng php sinh hc. S dng phng php vt l v ha hc gp nhiu tn km v t hiu qu hn phng php sinh hc. Nhiu ni, cc h ao b nhim, ngi ta tin hnh nui To, vi sinh vt, nh to tch ly cc kim loi, lm bin i thnh dng bay hi hoc kt ta xung, hoc cc cht thi l ngun thc n ca chng. C s khoa hc ca gii php ny l nhiu vi sinh vt c kh nng tch ly kim loi vi nng cao v trong t bo vi sinh vt mt s loi c nhng cht c bit gn vi cc kim loi. Cu trc c bit gn vi kim loi l nhng kim loi Fe, Mg, Cu, Zn, Mo, v cng c th tch ly nhng cht khc. Nng tch ly kim loi ca cc vi sinh vt thng cao hn mi trng bn ngoi hng ngn ln. Mi quan h gia sinh vt v tch ly kim loi c th hin qua m hnh sau:
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Hp th b mt
T bo vi sinh vt Tch ly bn trong
Bay hi (CH3)3Hg
Ta xung Fe, Mn, Mo, Ni Hnh VI.1. Mi quan h gia sinh vt v tch ly kim loi Ngoi cc bin php trn, ngi ta cn to cht bay hi: Methyl ha cc ha cht cha cc cht c: arsenic, telure, selence, thy ngn,to thnh cc cht bay hi. Ta xung: kh sulphate to thnh sulphite bay hi: H2SO4 +CH2O H2S + H2O + 2CO2 Nhng phn gn vi n trong ha cht s c ta xung. Hp th b mt: nhiu kim loi c t bo hp th b mt ca n nh to Chlorella hp th Au, mt s nm men hp th uranium trn b mt. Mt s vi sinh vt khc hp th Fe, Mo, Micrococcus hp th ch. Cc vi khun hp th b mt nhanh hn nm men. C nhng vi khun hp th cng h hng nhng cng c nhng vi khun hp th h hng ca chng xa nhau. Mt s vi khun hp th mt s kim loi c bit nh Bacillus megaterium 20oC gn 43 mg Cd/1g sinh khi. Tch ly t bn trong: sau khi hp th b mt, vi sinh vt tip tc a kim loi vo bn trong nh l thc n ca chng. Nu tch kim loi ra khi t bo th vi sinh vt b cht. Mt s vi sinh vt c th ht kim loi vo trong t bo vi nng gp hng triu ln so vi nng bn ngoi. So vi cc bin php khc nh vt l ha hc th bin php sinh hc c nhiu u th hn nh khng gy nhim, c th x l v thu hi nng thp. Vi sinh vt s dng cht hu c thi ra lm tng sinh khi ca chng, ng thi loi b c cht bn trong mi trng hoc c th x l mt cch hu hiu i vi tng kim loi thi ra mi trng. Bin php dng sinh vt x l cht thi mi trng hin nay ang tp trung nghin cu v ng dng (s cp n chng X, phn bn).
2. ng dng CNSH vo cng nghip khai khong.

CNSH khong cht (mineral biotechnology) - luyn kim bng con ng sinh hc (bihydrometallurgy) l vn mi mc du cch y hn
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1.000 nm, ngi La M bit ly ng t dng nc chy qua m ng. Vo th k XVIII, ngi Anh cng ly ng t nc chy qua m ng. Th k XVIII, ngi Ty Ban Nha (Spain) ly ng t m RioTinto (Nam M) nh mt s sinh vt tc ng vo khong sn, t con ngi thu c kim loi. Nhng trong mt thi gian di, con ngi t ch n hng ny. Dn dn tri thc ca con ngi ngy cng c tch ly, ngi ta c hng s dng cc vi sinh vt bin i cc khong vt tin hnh khai thc khong sn. Hin nay, s dng sinh vt khai thc ng, uranium, vng, bc, v nhiu kim loi khc. Gn y, ngi ta pht hin thy loi to Chlorella c kh nng hp th vng trn b mt t bo. Ngi ta tuyn chn ging to c kh nng hp th vng trn b mt ln. T m ra trong tng lai mt hng tm cc sinh vt c kh nng hp th cc kim loi qu him khai thc. Vic khai thc khong bng con ng sinh hc c u im l: - t tn nng lng - t lm nhim mi trng, t lm thay i cnh quan - C tnh c hiu cao, do mt s loi sinh vt hp th v bin i mt s nguyn t nht nh. 2.1. Tch qung bng vi sinh vt Nm 1947, t vi khun Thiobacillus ferroxydans c kh nng oxyha st, kh cc cht c lu hunh, v vy lm tan ng vo dung dch. Vo nhng nm 1950-60, ngi ta phn lp c cc ging vi khun ny. c im ca chng l sng trong mi trng c pH thp. Tip theo, ngi ta phn lp c mt s loi khc nh Leptospirillum ferroxydans sng pH=1-2. Khc vi loi trn l chng oxyha st, khng nh hng n lu hunh. Sau , ngi ta li pht hin c hng lot cc loi khc, c loi chu nhit cao nh Sulfolobolus bricleyi sng nhit 60oC bin i cc sn phm cha lu hunh. Ni chung, cc vi sinh vt bin i khong cht thng sng trong cc iu kin khc nghit m cc loi sinh vt khc khng sng c nh pH thp (1-3), chu ng c nng cht c cao m nng cc sinh vt khc b ng c nh arsenic nng 20 g/l, Cu: 60 g/l, Zn: 130 g/l. V c ch tc ng, chng c tc ng trc tip hoc gin tip cc phn ng sau: 4FeSO4 + O2 + 2H2SO4 2Fe2(SO4)3 + H2O
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hoc oxyha lu hunh: S2 + 3O2 + 2H2O 2H2SO4 + 283,6 kcal. Cn c th thc hin oxyha-kh theo phng trnh: 4FeS2 + 15O2 + 2H2O 2Fe2(SO4)3 + 2H2SO4 oxyha sphalerite (qung c cha km) ZnS + 2O2 ZnSO4 C th tc ng gin tip nh oxyha qung Cu (halkonite) Cu2S + 2Fe2(SO4)3 2CuSO4 (tan) + 4Fe2SO4 + S Qung uranite: UO2+ Fe2(SO4)3 (UO)2SO4 (tan) + 2FeSO4 + 1/2O2 2.2. ng dng vo cng nghip khai khong Polysaccharide do vi khun to ra c cu to v c tnh ha l khc vi polysaccharide ng vt v thc vt ch chng c nht cao khi nng thp. Do vy, chng c th c s dng mt cch tuyt diu lm cht gy nh tng, cht n nh dung dch nht, cht to ng c, cht n, T cc polysaccharide do vi sinh vt to ra c th dng nh mt loi du bi trn lm nh bt qu trnh khoan ging tm du m v kh t. Cc polysaccharide khc ca vi sinh vt cng nh nhng loi vi khun xc nh, nu bm dung dch hoc huyn ph ca chng vo ging khoan s to ra trong va qung s hnh thnh tch cc cc kh, p sut tt nhin s tng ln v hiu sut khai thc du s tng ln. Cht bo nhn c khi chit rt hydrocarbon t nm men nui cy trn parafin lng v nhin liu diesene li l tc nhn tt trong qu trnh lm giu khong sn bng phng php tuyn ni. Ti nguyn thin nhin khai thc nhiu lc b hao ht mt cch ng k, c bit l cc ti nguyn qu di t. l do k thut khai thc v ch bin chng cn lc hu. Lin hip x nghip khai thc luyn kim Dreskasgan ti 1/5, mt vi va ti 1/3 s lng qung ng cn st li trong t. Cc nh my luyn ch v cc nh my luyn kim khc tch li ti hng triu tn x. Trong cc x ny c cha ti hng nghn tn kim loi mu v kim loi him. Trong khi , dch huyn ph ca cc chng vi sinh vt c la chn li c kh nng ha tan ng, cobalt v nhiu thnh phn khc ca cc qung ngho v b thi. Cc dung dch c ly t t ln v thu thp li trong cc thit b x l bng ln men, s l cc ngun kim loi tinh khit c c m khng cn cc qu trnh luyn kim. Ngh nghip ca cc vi sinh vt luyn kim l rt khc nhau. Chng hn, nh vi khun c th tch c arsen ra khi dch c thic-ng-arsen, phn tch c dch c c ng-km v mt s dch c khc, tch c vng t qung arsenopyrite, thm ch tch c uranium t nc bin.
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Chc l trong tng lai s c nhng m t ng v cc x nghip khc nhm khai thc, lm giu, tinh ch luyn kim mu, kim loi him trn c s CNSH v c th hon ton khng c cng nhn. Cc sn phm ca vic sn xut nh vi sinh vt t ra c li c i vi cc nh luyn kim en, cc x nghip sn xut t, xe tng. Cc dn xut lipid vi sinh vt, cc monocarbonic ca cc acid bo c th l du bi trn tuyt diu i vi cc trc my cn khi sn xut cc tm thp mng, cn phospholipid - cht ph gia chng n mn c a vo du nhn. T furfinol ca cng nghip thy phn, ngi ta sn xut c nha feran v furil tng hp. l nhng cht kt dnh tuyt vi dng sn xut cc ct cao cp. Cc ct ny rt cn thit cho cc l thp, gang, ng v nhiu chi tit khc c hnh dng phc tp, cn cho cc khun kim loi c cht lng cao, cho php gim bt hoc loi b hn cc x l c hc tip tc v lm tit kim c kim loi. Hin nay trn th gii c nhiu nc s dng CNSH tch cc kim loi him qui m ln. Ngoi vic tch ng nh vi khun Thiobacillus perroxyd thu ng gp 5-6 ln so vi khai thc thng thng. Ngi ta : - Tch uranium: Canada c n 75% uranium dng oxyde U3O8. M Denison dng vi sinh vt khai thc. Nm 1986, khai thc t 360.000 kg U3O8. Trong tng lai, Canada s m hng chc x nghip sn xut qung uranium. - Tch vng, bc: Au, Ag thng ln trong phc cht pyrite FeS2. Thng pyrite bao ngoi cc cht . Lc u M, Australia khai thc vng bc pyrite ngoi rt nhiu kh khn. Khai thc khng hiu qu. Sau ny Lin X (c), M dng vi sinh vt lm tan pyrite ngoi, l vng ra. Nh vy m vn u t th t m hiu qu kinh t th cao. TI LIU THAM KHO Phm Thnh H, 1994. Cng ngh sinh hc. Ti liu nh my trng i hc Khoa hc T nhin thnh ph H Ch Minh. Nguyn B Lc, 1999. Gio trnh Cng ngh sinh hc. Nxb Gio dc H Ni. Trng Vn Lung, 1995. Cng ngh sinh hc. T sch i hc Khoa hc Hu. Nguyn Vn Uyn, Nguyn Tin Thng, 1996. Nhng kin thc c bn v cng ngh sinh hc, Nxb Gio dc H Ni.
1. 2. 3. 4.
CNSH ch bin sn phm thu hoch
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Chng VII
Cng ngh sinh hc ng dng trong ch bin cc sn phm nng lm ng nghip

Lch s ca ngnh hc ch bin chuyn nghin cu v nhng bin i sinh ha ca ng cc c pht trin theo yu cu ca cng ngh bo qun v ch bin lng thc. Phn ln cc cng trnh u hng vo nghin cu cc thnh phn ha hc ca ng cc (go, bt, m si, bnh m,). Ngi ta nghin cu nhng cng ngh c truyn v cng ngh mi ch bin cc sn phm ph hp vi nhu cu, tp qun sng ca con ngi. Sau y l mt s v d.
1. Cng ngh ch bin c truyn

1.1. Sn phm cng ngh ch bin t glucid (saccharide) thc vt T xa ng cha ta bit ch bin cc sn phm t tinh bt cho chng va khi b h hng va tng cht lng v hng v ca sn phm. Nguyn liu ch bin tinh bt l cc loi c, ht nh ng, khoai, sn. Hm lng ng bt rt cao (>70%). Ngi ta ch bin phn ln bng phng php th cng hay my mc. Cc dng tinh bt thng c ln men hoc chua, sau ty theo sn phm cn dng m c nhng phng thc ch bin khc nhau c hng v khc nhau. Ph bin trong dn gian, ngi ta lm bnh n hng ngy, lm ko, lm ng v cao hn na l lm ru. cc nc chu u c ru nho ngi ta lm bng con ng th cng truyn thng xut hin hng ngn nm nay. cc nc trong khu vc ng Nam cng c cc loi cm ru nh cm ru np Indonesia, vang qu Philippines, ru sak Nht Bn. Vit Nam c ru du ty, ru cuc li, ru np than, ru vang , vang trng, ru np (ru ), ru mu (ch bin t hoa qu) v.v. 1.2. Sn phm cng ngh ch bin t protein ng vt v ch bin ln mem t thc vt Ngoi glucid ra, protein l thc phm khng th thiu c i vi con ngi. Thm ch, cc nc pht trin protein l ngun thc phm chnh. S ngho i l s thiu protein. Ni cch khc, ngi ta ly tiu ch protein cung cp nhiu hay t i vi con ngi nh gi nc thuc vo din ngho i mc no.
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Protein cung cp cho con ngi ly t nguyn liu ca ng vt, thc vt thm ch t vi sinh vt. Trong dn gian, tht, tm, c l mn n hng ngy c cha protein. Di tc ng ca iu kin bn ngoi nh nhit , m, vi sinh vt ln men thi, protein d b h hng. V vy, t xa ngi ta c nhng phng php bo qun nh x l bng nhit : phi nng, hun khi, nng, quay, sy bng h thng nng, sy thng hoa, lm lnh (p lnh, lnh ng); bo qun bng phng php ha hc: p mui, p bng acid, cht khng sinh, v.v. Ngi cng tin hnh ch bin, chuyn ha tht, c c sn phm ngon ming hn nh lm mm, lm tm chua, nem chua. Mt s a phng c nhng ch bin truyn thng nh lm mm nm, mm c thu, nc mm Phan Thit, nc mm Ph Quc, tm chua Hu, nem chua Thanh Ha, nem chua Hu, Tm mui thnh Balao Balao (Philippines), tm mui Kung Chom (Thi Lan), c thu mui Kysaya, mc mui Ika Shiokara (Nht Bn); v.v. V thc vt ngi ta ch bin thnh cc sn phm giu protein nh lm tng, chao, nc chm, x du, u h, sa u nnh ti, v.v. Trong cc loi thc phm ln men truyn thng th miso ca Nht Bn (700.000 tn/nm u thp k 60 ca th k trc) rt ng ch . Dng u tng nguyn ht v ln men nh Aspergillus oryzae. Nht Bn dng miso 24 g/u ngi/ngy. u ph, chao, tng Trung Quc (sufu) v c Vit Nam l sn phm ging nh fromage (phomat) c ch bin nh nm mc (ch yu l Mucor). Ang-kak l mt loi thc phm ch bin Trung Quc bng cch ln men go nh nm mc Monascus purpurcus. M v nhiu nc khc ngi ta a dy chuyn sn xut qui m ln cc sn phm ging nh tht nhng ng l khng h cha tht. Ngun protein y l u tng c c c li hoc tch ra. T dung dch ny c hoc khng b sung casein v albumin trng), ngi ta nhn c mn tht ng, dng k thut xe si t lm thnh th, to ra cc mn tht (gi, ch, tht rn, biptt, tht mui, tht bm, gim bng, tht th rng, v c tht nghin pat). Vit Nam nhng cha chin v c nhng Pht t theo o Pht, ngi ta cng dng cc mn n chay, nhn b ngoi l nhng mn n bng tht nhng khng phi t tht. 1.3. Sn phm cng ngh ch bin t cc loi du thc vt v m ng vt trong thc vt, lipid c tn l du, ng vt ngi ta thng hay gi l m. Lipid hay cht bo l nhng hp cht hu c c trong t
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bo sng, khng ha tan trong nc, nhng tan trong cc dung mi hu c, khng phn cc nh ester, ether du ha, toluen. Lipid l thnh phn cu to quan trng ca cc mng sinh hc l ngun nguyn liu d tr, cung cp nng lng cho c th. Lipid thng c chia thnh hai nhm ln: Lipid n gin l ester ca alcohol v acid bo. Thuc nhm ny c triaxylglycerol (du thc vt, m), sp v sterid. Lipid phc tp: trong phn t ca chng ngoi alcohol v acid bo cn c cc thnh phn khc nh cc gc acid phosphoric, cholin, saccharide. Nhm ny c chia thnh cc nhm nh ty theo bn cht v cu t alchohol hoc cu to khc nh glycerophospholipid, glyceroglycolipid, sphingophospholipid v sphingoglycolipid. Phn lipid ca thc phm, cc thc phm ch bin ch yu t lipid ng thc vt, m, du bo tinh luyn u l nhng i tng cho vi sinh vt gy h hng. So vi thc phm khc th m v du bo tinh luyn t b h hng bi vi sinh vt hn. Chnh cc sn phm giu lipid ny h hng v ha hc nhiu hn v vi sinh vt. Hnh thc h hng chnh ca lipid l qu trnh thy phn v oxyha hoc phi hp c hai qu trnh ny. Kt qua lipid b chuyn ha thnh glycerine, kh CO2 v nc. Biu hin h hng ch yu ca cc loi m v du bo tinh luyn l hin tng i, kht. Cc sn phm lipid b i thng c mi v rt kh chu. Trong qu trnh i, cc phn ng thy phn v oxyha thng xy ra ng thi. Vic ch bin du m c truyn ca nhn dn t trc n nay l p (i vi thc vt) v rn (i vi ng vt). V d: Lc c cha nhiu du v protein, l loi thc vt qu. Lc c dng lm du n, b nhn to. Kh lc c dng ch bin nc chm, ch phm protein, bnh ko, thc n gia sc, v.v. Bn thn ht lc dng lm thc n, lc rang. Qu lc c lp v cng, c tc dng bo v rt tt cho ht trnh c nh hng xu ca mi trng bn ngoi. Cc gia nh nng thn c th dng 2 lp ct quy quanh, gia l lp tru kh hoc tro bp chng m. Nu s lng t c th bo qun bng chum, vi v bt kn. Trong ch bin c truyn, th cng, ngi ta hay p dng phng php p. Lc sau khi c la chn, em vo nghin nt. Mc ch ca qu trnh ny l ph hy trit nhng t bo nhn nhm gii phng du ra dng t do d thu hi. m thch hp cho qu trnh nghin l 1012oC. Bt nghin c a qua chng sy lm cho du c linh ng v khi p du d dng thot ra.
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Ngoi lc ra, ngi ta cng s dng nhiu thc vt khc p du nh c du, da, o ln ht (iu), liu, i hi (m ln, mc p mua), hng dng, bng, v.v. ty theo nhu cu s dng ca chng. X du (shoyu) ch Trung Quc cch y nhiu th k v sau lan ra cc nc khc Vin ng, Nht Bn l nc sn xut ch yu hin nay. Ch to x du nh ln men ca Aspergillus oryzae. V m ng vt th ngi ta thng hay rn nht l m ln, m b, m cu, m g, vt, m c, v.v. Ni chung, m c nng lng cao cung cp cho nhu cu sng nn ngi s dng kh ph bin nht l cc nc vng n i, hn i. Tuy nhin, trong m c nhiu cholesterol gy x cng ng mch, nn sau ny nhiu nc s dng ch yu l du thc vt. 1.4. Sn phm ch bin t rau qu Rau qu l mt trong nhng sn phm c t l tht thot cao nht, c khi ln ti 100%. Mt trong nhng yu cu quan trng nht i vi vic ch bin v bo qun rau qu l lm th no bo tn mt cch ti a cc cht dinh dng, hng v, mu sc ca rau qu. Mun vy, cn phi hiu r thnh phn ha hc, cc qu trnh ha sinh xy ra trong rau qu. Nhng tnh cht quan trng nht ca qu nh hng v, mu sc, bn khi bo qun u c hnh thnh trong qu trnh chn. Vic nghin cu qu trnh sinh l, ha sinh c bn ca s chn c ngha to ln trong vic tng cng phm cht ca qu. Xc lp thi hn ti u trong thu hoch v ngn nga nhng tn tht khi bo qun v ch bin. l nhng qu trnh sinh trng ca qu khi chn, nhng bin i v thnh phn ha hc ca qu khi chn. Nhn dn ta t xa n nay, qua qu trnh sng tch ly nhiu kinh nghim, nn thu hoch qu vo lc no dng vo mc ch g. T nhiu th k nay, con ngi bit bo qun v ch bin rau qu, l loi thc phm nhanh b h hng. Trc ht ngi ta ch n vic chn lc cc ging cy cho qu v rau c cht lng tt v bo qun lu. Tip theo l lai hu tnh, ng thi kt hp vi bin php chm sc cy nhm mc ch to ra cc ging rau qu c cht lng mong mun. Rau qu th c nhng loi cn bo qun tt s dng, khng cn qua ch bin, c loi th cn phi tin hnh vic ch bin hoc chuyn ha sn phm. V d: cam, bi, cn bo qun bng cch vi trong ct hoc cho vo chum nc ng, nc mt va dit khun va khng c khng kh, gim ti a qu trnh h hp: kh CO2 v nc khng thot ra lm qu ti lu. Rau qu cho vo t lnh chng s ln men thi. Mt s sn phm khc nh hnh, kiu, mng th phi nng. C loi th mui chua nh da chut, cc loi ci, c chua, v.v. C loi ch bin thnh ru
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nh nho, du, m, mn, v.v. Qua s bo qun v ch bin phong ph ny khng nhng sn phm khng b tht thot m cn lm tng hng v thm ngon hoc cht lng tt hn c khi dng ti sng.
2. Mt s qui trnh cng ngh ln men hin i

Mc du CNSH ch yu c thc hin v pht trin cc nc ang pht trin vi cc thnh tu nh ln men lin tc - bt ng ha enzyme v t bo, cng ngh di truyn, v tnh ha cc gene (gene cloning), cc b quyt v cng ngh c truyn cng khng h thiu ti cc nc ang pht trin. Tuy vy, cc nh nghin cu Nht Bn vin Nghin cu Vt l v Ha hc (RIKEN, Bo tng ging vi sinh vt Nht Bn), vin Nghin cu Vi sinh vt hc ng dng thuc i hc Tng hp Tokyo v b mn CNSH thuc vin Nghin cu Khoa hc v K thut Thi Lan (b Khoa hc v K thut Bangkok ( phn lp c 80 chng nm men t 39 mu thuc 29 loi thc phm khc nhau (c, tm, tht, ng cc, hoa qu,) v cc vt liu c lin quan khc Thi Lan. Cc chng ny thuc v cc chi Candida, Debaryomyces, Geotrichum, Pichia, Saccharomyces, Saccharomycopsis v Stephanoascus. Cc nghin cu ny c tin hnh nhm mc ch lm sng t vai tr ca cc loi nm men trong qu trnh ln men v lm chn cc thc phm ln men lm tng cht lng ca cc sn phm ln men.Cc qu trnh ny chu s nh hng ca s hin din nhiu loi vi sinh vt khc nhau. Chng hn, vi khun lactic v nm men ng vai tr quan trng trong vic sn xut v lm chn ru sak, nc chm, tng, fromage (phomt) v.v. S phi hp tc ng ca vi sinh vt cng ni bt trong cc qu trnh sn xut v lm chn thnh thc nhiu loi thc phm ln men vng ng Nam (Suzuki M. et al. 1987). Nhng mt di y cho thy r nhng li ch m cc nc ang pht trin c th t c trong ng dng CNSH hin i vo vic ch bin cc sn phm nng lm ng nghip da trn nhng b quyt ca CNSH c truyn v CNSH mi p ng nhu cu v thc phm v dinh dng. 2.1. Cng ngh ln men t nguyn liu giu glucid Trong qu trnh ch bin, CNSH ngy cng pht trin ng gp phn ng k ch bin cc sn phm c cht lng cao. Ln men trn mi trng xp (mi trng bn lng) Trong cc iu kin ln men nhm nng cao gi tr dinh dng v gi tr kinh t cho nng sn v ph phm ca cng nghip thc phm th ln men trn mi trng xp l nhng phng php cng ngh rt ng quan tm.
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Indonesia, Tempeh cha n 55% protein l loi bnh xp lm bng u tng (bnh Kedeke), bng lc (bnh Ontjom) hay bng da (bnh Bongkrek). Ch to Tempeh cn 2-3 ngy. u tng c ngm nc 12 h, bc v, luc u trong 30 pht ph cht c ch enzyme tripsine ca ty tng v mt cht c ch sinh trng khc. Ra li u nhiu ln v ra nc. Cy bo t nm Rhizopus oligosporus v cc ht u c nghin nh ng trong khay, s ln men ko di 36-38 h nhit 34oC. Thu c bnh mu nu nht. Tempeh c n sau khi nu hay rn vi du da. Khi ln men s xy ra qu trnh thy phn cc thnh phn glucoside cng nh 30% triglyceride, lm gii phng ra loi acid bo ch yu l acid linoleic (2,5 g/100g) trong khi hm lng lipid trong Tempeh vn gn bng lng lipid trong u tng (20-26%). Lng protein tng ln v t ti 50-55% trng lng kh (u tng cha 40-43% protein) v phn ln protein b enzyme protease ca nm mc thy phn thnh cid amin nn lng cha nitrogen ha tan tng t 0,5-2%. Nh vy m lm thay i cht lng ca u tng khi dng Tempeh. Tempeh cng kh giu riboflavin (vit. B2: 7g/g) v niaxin (vit.PP: 60g/g) (vit. B2 cao hn 2 ln v vit. PP cao hn 5 ln so vi u tng). Tempeh cn cha vit. B 12 (5-30 mg/g). Cc nh khoa hc M ng dng cch ch bin Tempeh ca Indonesia vo vic ch bin ng cc v Sn. Bt M c ln men bng Rhizopus oligosporus nhit 31oC trong 20 h c cha lng protein cao hn ban u 6-7 ln, riboflavin cao hn 5 ln v niaxin cao hn 2 ln. R. oligosporus to ra t amylase, do bt M khng b thy phn v khng xy ra qu trnh ln men ru. Raimbault (Php) nghin cu Senegal s thy phn bt sn bng vi khun Lactobacillus nhm bo qun sn ti hoc cy thm nm si lm giu protein cho bt sn. Cc chng vi khun Lactobacillus c kh nng thy phn tinh bt c phn lp t sn v c dng ln men lactic nhm ci thin iu kin bo qun cng nh nng cao gi tr dinh dng cho bt sn. Mexico, Raimbault tch c 100 chng vi khun lactic t cc thc phm ln men truyn thng nh pulque, pozol, tesguino. Nhng chng ny thuc v cc chi Lactobacillus, Pediococcus v Streptococcus. Mt chng Lactobacillus c la chn dng vo vic bo qun sn ti v lm giu protein cho sn. Sau khi thc nghim ln men phng th nghim, ngi ta trin khai ra cc qui m sn xut th, nh gi cht lng dinh dng ca cc
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sn phm ln men, nh gi nguy c nhim cc chng t bin sinh c t nm (mycotoxin). Ngoi Mexico, cc nc khc Nam M (Argentina, Chil, Venezuella) cng cc nc Trung M v vng Carib (Costa Rica, Cuba, Guatemala) cng quan tm n cc ng dng ln men trn mi trng xp lm giu protein cho cc c cht giu tinh bt to nn cc thc phm ln men giu protein gi tt l AFEP, lm ln men lactic nhm bo qun cc sn phm nng nghip giu tinh bt, lm tng kh nng tiu ha v tng hng v cho cc thc n chn nui (b ma, thn cy la min), gia sc ln, tm ra cc enzyme phn hy tinh bt, phn hy cellulose mt cch r tin, kh cc cht c hi ra khi cc nng sn phm (kh tannin thuc loi polyphenol v cafein ra khi ht c ph). Ngoi ra, ngi ta lm giu protein cho mt s loi sn phm m trc y ngi ta phi chn hoc th xung bin hng triu tn, nay bin chng thnh thc n cho chn nui gia sc, tm, c hoc lm phn bn hu c vi sinh. Mc d vic sn xut cc thc phm ln men giu protein t glucid cn c nhiu kh khn v k thut v v kinh t, nhng y l mt lnh vc y trin vng trong vic t cung t cp thc n v nng cao gi tr dinh dng cho cc khu vc dn c thiu ht protein. Hn na, chng cn y trin vng trong vic sn xut cc sn phm sinh ha hc c gi tr cao nh enzyme. Ch bin glucose t g Ngh nghip v g rt a dng. Theo tnh ton ca cc chuyn gia, cch y 40 nm, cc mt hng ch to t g gm khong 4.000 loi, by gi qu 25.000 loi v cn c tip tc m rng. Nhng bt hnh ch, trong qu trnh x l c hc, rt nhiu g b hao ph mt cch v ch. Ph phm ca vic ch bin g trn l 27% khi ca g, trong cng nghip g dn: 55%, trong sn xut dim: 65%. Khi ch bin theo kiu c ha hc, cc mnh vn ca g hoc cc ph liu s c tm nha ri c p li v nh m ch to ra cc tm g p-v bo, g p-si dng thay th cho g nguyn vn. Ch bin ha hc g l c hiu qu. Cc nh my cng nghip ha hc rng x l g nhit cao v sn xut ra acid acetic, than g, du thng, methanol, formalin, phenol, aceton, v hng lot cc sn phm khc cn ch xut t gc cy thng s thu c caniphol. Trong cng nghip cellulose-giy, trc ht t g phi tch c cellulose, sau t cellulose mi lm ra giy. Cellulose cn l nguyn liu sn xut ra si nhn to visco, cellophan, phim nh, verni v nhiu sn phm khc. Cellulose l mt trong nhng cht hu c n nh nht, c thc vt to ra nh nng
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lng nh sng mt tri. Khng c loi thc vt no, khng c loi ng vt no (tr loi nhai li c cc loi vi sinh vt trong d dy sinh ra enzyme cellulase) c th phn gii v ng ha c cellulose. Cellulose to ra b xng chng cho cy tho v thn cy g. Trong cc mng t bo ha g, cc vi si (microfibril), cellulose ng vai tr ct sc ca b tng, cn ci ng b tng y l lignin 2030% v hemicellulose 20%. Khi sn xut cellulose th lignin v cc thnh phn hemicellulose cn li s c ha tan bng cc mui ca acid sulphuric hoc bng cc cht kim. y khng xy ra nhng chuyn bin ng k i vi cellulose. cc nh my thy phn th phng hng ca qui trnh cng ngh li hon ton ngc li: cellulose v hemicellulose chuyn vo dung dch, cn lignin th li dng cht rn t nhin. S khc nhau cn bn li ch khc. S thy phn khng phi c mc ch duy tr m l cu to li hon ton phn t cellulose. Thm ch, c th ni rng, y hon ton vic tr ha li phn t ny. Khi quang hp trong l cy to thnh glucose, hng nghn phn t glucose ngng t li, polymer ha thnh phn t polysaccharide-cellulose v khi gii phng ra nc. Khi thy phn, phn t cellulose lin kt vi nc phn gii ra hng nghn gc glucose cu to ra n. Cellulose l vt cht c bn ca g, hon ton khng n c, li chuyn ha thnh glucose - loi hydrate carbon sinh ra trong cc l xanh khi hp th nh sng mt tri. Loi ng ny s ha tan trong nc v d dng c tt c cc c th sng ng ha. Vic thy phn g v cc vt liu thc vt khc nh v ht bng, v ht hng dng, thn v b ng, rm r ca la v cc loi ha tho khc l mt qu trnh ha hc kh phc tp. Trong t bo thc vt to thnh cc polysaccharide phc tp (cellulose v hemicellulose) t cc ng n gin nh nng lng nh sng mt tri. Khi phn gii ngc li cng cn dng mt nng lng tng t. Mc d qu trnh thy phn tin hnh ch c mt ca cc cht xc tc (acid sulphuric, chlohydric hay cc acid khc), khi cht phn ng (g) nghin nh cng vi cht xc tc phi un si n 180oC hay cao hn, vi mt p lc trong thit b thy phn n 10-12 atm.. Dung dch ng acid to thnh c lin tc ly ra khi ni phn ng, cn phn rn l lignin th sau khi kt thc ton b qu trnh, s ly ra khi thit b thy phn. Dch thy phn c trung ha v lm sch cc to cht. Dung dch tinh khit c lm ngui (dch trung ha neutralizate) cha khong 3% ng hexose (glucose, mannose, galactose) v pentose
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(xylose, arabinose). Trong dung dch cn mt s lng acid acetic v acid hu c khc. cc nh my ru thy phn nh mt loi enzyme bin ng hexose thnh ethanol, b cn li, sau khi ct ru li c dng nui cy nm men gia sc. Dch trung ha, sau khi loi bt nc s l mt dch m c cha 25-30% ng gi l mt thy phn, dng trong khu phn ng vt khi thiu hydrate d tiu. Vic nghin cu a vo sn xut qui trnh cng ngh mi tin b hn, cho php thu c dch ng nhanh hn, r hn, v sn xut glucose thc phm, nm men bnh m, nm men thc phm, acid amin khng thay th (lysine), acid glutamic, acid citric v nhiu sn phm c ch khc. Ch bin rm r Cc ch phm enzyme to ra kh nng nng cao cht lng thc n ti v s dng trit hn cc sn phm ph ca trng trt nh rm r, chng hn, lm thc n cho chn nui. Trn ng rung, ng c, ngi ta gieo trng ngy cng nhiu cc thc n xanh dng trong chn nui. iu cn thit l phi bit ch bin thc n v bo qun dng quanh nm m khng b mt dinh dng Phc v cho mc ch ny, ngi ta chua, bo qun v ma ng thc n chn nui bng acid lactic sinh ra bi vi khun lactic Khi chua cc cht dinh dng trong c vitamin s b mt mt t hn so vi c kh. cho vi khun lactic pht trin nhanh trong khi lng thc n xanh bng cch nn k v y kn, cn c lng ng ha tan v d ln men. Trong thc vt cng cha nhiu protein th cc vi khun ny cng cn c nhiu ng hnh thnh acid lactic v mt phn acid acetic. Cc acid ny s lm acid ha khi thc n chua v lm c ch s pht trin ca cc vi khun butyric trong loi vi khun ny lm thi ra khi cht xanh. Khng phi mi thc vt u c cha lng ng cn thit cho s pht trin ca vi khun lactic v cho vic lm chua mt cch thch hp khi lng thc n chua. Cng v thiu ng m cc cy trng giu protein nh u Ha Lan, c Ba l, Du tm, kh chua, cn c inh lng v u tng thuc loi thc vt khng chua c. S phn loi nh vy nhm a thm ch phm enzyme vo hp l chuyn cy h u thnh loi chua c. Mun vy, khi xp thc vt vo ni ct cha, phi cho thm mt lng khng ln cc ch phm enzyme. Di tc dng ca chng, mt phn polysaccharide (cellulose, tinh bt, hemicellulose, pectin) b phn gii thnh ng v cc acid hu c n gin, tan trong nc thnh acid amin. Tt c cc cht ny c vi
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khun lactic ng ha tt v sinh ra acid lactic mt cch nhanh chng, thc n xanh nhn c s c cht lng tt, ti, ngon v phong ph cht dinh dng. B sa, b c, b, ln u n ngon lnh. Thc n chua nh vy bo qun c 2 nm. Nh cc ch phm enzyme c th chuyn ha rm r thnh thc n chua c cht lng tt, thm ch thnh mt loi thc n glucid-protein qu gi. Cc chuyn gia ca vin Nghin cu K thut Sinh hc Lin X (c) v vin Hn lm Nng nghip mang tn Timiriazev K.A. ngh mt phng php n gin, r tin v c hiu qu x l rm r bng nhit v enzyme trong cc my trn hn hp C-12 do ngnh cng nghip Lin X sn xut. Enzyme chuyn ha phn polysaccharide phc tp ca rm r thnh cc dng ha tan v d c vi sinh vt ng ha. Sau c chua hoc c cy nm men pht trin trn phn ng ca thc n. Trong trng hp ny thc n t rm r c cht lng dinh dng gn vi u. Ch bin tinh bt ng Sau y l mt v d v cng ngh ch bin ng ca nh my sn xut tinh bt ng v tinh bt ng ci bin ca cng ti Hu hn Khai thc ng Tn Nguyn, Ct Lm, Trung Quc (lin doanh vi nc ngoi. Vn ng k kinh doanh 12 triu USD. Tng kinh ph u t l 45 triu USD). Sn phm ch yu ca cng ti ny l tinh bt. * Tinh bt ng c th dng lm nguyn liu trong cc ngnh cng nghip khc nhau: lm giy, dc liu, dt, thc phm. - ng dng trong cng nghip thc phm: Tinh bt ng c th chuyn ha thnh ng. V d: fructose t tinh bt ng c th ci thin v tng cng khu v ca cc thc phm ch bin. Ngy nay, ng tinh bt (starch sugar) l ngun ng mi dng thay th ng ma (ngt t 173,3% so vi ng ma). Tinh bt ng cng l nguyn liu sn xut m chnh, acid citric, v.v. - ng dng trong cng nghip dc: Tinh bt ng c th sn xut ng glucose dng trong y hc nh qu trnh thy phn. Sau , glucose c th sn xut vitamin C v.v. Tinh bt ng lm cht ph gia trong vic sn xut thuc vin v lm nguyn liu ch bin cc tc nhn chng vi khun, chng nm v.v. - ng dng trong cng nghip giy: Tinh bt ng lm cht ph gia trong cng on t ca qu trnh sn xut giy khin cho giy bn chc v c b mt p. C th lm cht h dnh b mt tng bng ca giy, sn xut ra b mt giy in tng
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i tt, gim bt t l rch v x xc. C th lm cht kt dnh cho lp bi nguyn liu mu gia giy v ba cng. Tinh bt ng lm cho mu kt dnh vi nhau v kt dnh vi si to thnh b mt bn chc, trn u n v lin tc, lm tng trng, sng v trong sut ca giy. C th lm cht kt dnh sn xut giy cng, giy carton v.v. - ng dng trong cc lnh vc cng nghip khc: Tinh bt ng dng trong h si trong cng nghip dt, lm cht ph gia trong cng nghip c, trong cng nghip khoan du cng nh trong cng nghip cht do. * Bt gluten t ng: Bt gluten t ng l mt loi bt giu protein (>60%), cha nhiu acid amin chng c gi tr cao trong ch bin thc phm v gia v. Loi bt ny c th dng lm mi trng nui cy trong cng nghip ln men. Thc n gia sc lm bng bt gluten l thc n cao cp c cht lng tt i vi gia sc, gia cm, c. * Du phi ng: Du phi ng c cha nhiu acid linoleic c th ha tan cholesterol trong mu, lm mm huyt qun, lm gim huyt p v ngn nga hin tng x va ng mch. y cng l loi du thc phm c cht lng cao, cha nhiu vitamin E, l mt loi du b dng ho hng. Ngoi ra, du phi ng dng lm nguyn liu ha hc sn xut x phng v cc loi sn phm khc c lin quan n du m. * Ch phm c hm lng fructose cao: ch phm fructose l dung dch khng mu, trong sut. Cc cu t ca ch phm l fructose v dextrose. Ch phm hm lng fructose cao c mang danh hiu Mt Ong nhn to v hin nay thnh hnh khp th gii. Chng khng nhng c v thun khit, c ngt cao hn ng ma m cn c u vit khc nh gi tr dinh dng cao, d hp th. Nhng c tnh ca ch phm ny v tnh thm thu (permeability), v tnh d kt tinh (crystallization), v tnh ln men (fermentation) v tnh chng chu su rng v.v. u l nhng nguyn nhn ngi ta u tin cho ch phm ny trong cng nghip thc phm, trong cng nghip ung v trong vic tiu dng gia nh. c bit ch phm ny dng trong cc bnh nhn mc bnh i ng v trong ung ca cc vn ng vin. Ch phm c hm lng fructose cao ca cng ti Tn Nguyn c loi F-42 fructose (42% fructose), F-55 fructose tiu chun (55% fructose). * Tinh bt ng ci bin (corn modified starch): cng ti Tn Nguyn sn xut 5 loi tinh bt ng ci bin: ci bin nh acid (acid modified starch) bng cch cho tinh bt huyn ph trong dung dch acid long (HCl hoc H2SO4) nhit trong phng n nhit h ha. Khuy cho n
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khi nhit nht huyn ph gim n mc thch hp, trung ha bng kim hoc sodium carbonate, ra v sy kh. Loi tinh bt ny c s dng lm cht h si trong cng nghip dt, dng ch to cht ngt mm trong cng nghip thc phm, dng lm cht h dnh cc giy c bit trong cng nghip giy, dng trong cc hiu git l lm sch qun o, dng lm nguyn liu c bn sn xut cc cht ty ra dng lng trong gia nh, dng sn xut giy cng dn tng. Ci bin bng cch oxyha hoc cn gi l tinh bt b oxyha (oxydized starch), c oxyha nh mt s tc nhn oxyha nh natrium hypochloride (NaOCl), calcium hypochloride, ammonium persulphate, kalium persulphate, hydroperoxyde, acid peracetic, kalium permanganate, perborate, v.v. Phn ng c tin hnh trong dch tinh bt huyn ph hoc trong tinh bt nho c h ha. Ty thuc vo chng loi ca cht oxyha v ty thuc vo cc iu kin ca phn ng, nhm carboxyl (-COOH) v nhm carbonyl (-C=O) c gn vo tinh bt, ng thi xy ra s gii trng hp ha (depolymerization). S oxyha trong trng hp ny ch l mt qu trnh trong mt nhn ci bin (modification reagent) s gy ra hai bin i quan trng v mt ha hc (s gii trng hp ha + s gn cc nhm carboxyl vo tinh bt). Nhng tinh bt ny c dng trong cc tc nhn kt dnh trong cng nghip giy, h si trong cng nghip dt, dng trong cng nghip thc phm v dng lm cht kt dnh dn giy ba, giy p tng, ba cch m trong cng nghip vt liu xy dng. Tinh bt c lin kt ngang (cross linked starch) l kiu ci bin thu nhn c t hot ng ca cc nhn 2 chc hoc a chc (polyfunctional reagents) c kh nng phn ng vi nhiu nhm hydroxyl v t nht l 2 nhm hydroxyl. bng cch nh vy s hnh thnh lin kt ngang 1 phn t tinh bt ny n 1 phn t tinh bt khc. Tinh bt ny s dng trong cng nghip thc phm, trong gia v ca xp, nc tht, nc xt, v.v., trong ngh lm bnh pudding v cc loi thc phm khc c tht nhi, dng lm cht hp th ca cc dng v sinh ca ngi, lm vt liu hp th cc cht trao i ion, lm cc tc nhn chng ngng kt trong phim nh. Tinh bt cationic (cationic starch) l mt loi ester tinh bt c th iu ch theo phn ng sau:
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NaOH
St-OH + CH2-CH-CH2-N (CH3)3Cl O Tinhbt+ammoniumchloride 2,3epoxypropinmethyl St-OCH2-CH-CH2-H+(CH3)3 + 3ClOhydroxypropinmethyl tinh bt + ion chlore Loi tinh bt ny dng trong cng nghip giy, cng nghip xy dng, dt, cng nghip x l nc thi cng nh trong cng nghip khai khong. Ester tinh bt (starch ester): acetate tinh bt hoc tinh bt c acetate ha (starch acetate, acetylated starch). Acetate tinh bt c iu ch theo phn ng sau: O HC-C O St-OH + O NaOH St-O-C-CH3 + CH3COOH H3C-C O Tinh bt + aldehyde acetic NaOH acetate tinh bt + acid acetic Loi tinh bt ci bin ny c s dng trong cng nghip thc phm, dt, giy v dng lm tc nhn ht m trong cc ngnh cng nghip khc nhau. Quay li vn nghin cu sirofructose, bi v n l loi ng c ngha quan trng trong nhng nm ti y. Nm 1986, Usami trng i hc Waseda tng hp c mt cht gy v ngt mi, cht xylofructose. Loi ng i ny c tng hp nh enzyme bfructofuraloxydase ca nm Penicillum frequentem, enzyme ny xc tc thy phn saccharose thnh fructose v glucose, ng thi em mt gc xylose gn vi fructose. Hng Lotte Inc. hp tc sn xut xylofructose. Mt loi ng ba cng c tng hp t fructose l galactozilfructose nh s dng enzyme ca vi nm Aspergillus spp. do hng Meji Seaki Co. Lid. ca Nht Bn tin hnh. Theo mt s chuyn gia siro (xi-r) c nhiu ng fructose ch t ng (HFCS) sn xut bng ng phn ha enzyme glucose ch t tinh bt ng thy phn s thng tr trn th trng cht ngt nhiu calo t nhin trn th gii thp k 1990. Xir ny s cung cp 50% th trng th gii, trong khi cc cht ngt tng hp ng gp khong 10%.
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cc nc pht trin, tiu th sirofructose ang dn dn thay th ng saccharose. Th d M, tiu th ng saccharose gim t 91 pao/u ngi nm 1979 xung 87 pao/nm 1980. Trong khi tiu th sirofructose t 15,4 pao ln 19 pao/u ngi cng trong mt giai on . Nm 1987 tiu th ng ma v sirofructose theo u ngi M l tng ng 67 pao v 36 pao. S bnh trng th trng sirofructose v sn xut cht ngt ny l do tin sau y quyt nh: - Cc nc nhp khu ng pht trin mnh, sn xut sirofructose (isoglucose) hoc trin khai cc cht ngt thay th. - Cng nghip tiu th nhiu ng dng lng, mc d tiu th ph thuc vo tng ngnh cng nghip thc phm. - Gi ng cao, nh hng n cng nghip ln ngi tiu th (tr nhm dn c thu nhp thp). - Cc h thng chuyn ch v phn phi nng sut cao c th p ng nhu cu tn tr v s dng xi-r cng nh cc ph phm nh du phi ng v thc n gia sc t ng. - Cng nhn c trnh cao, cung cp nc sch, ha cht, enzyme v nng lng. - C vn, v xy dng x nghip isoglucose i hi vn u t ln. - C di do ngun tinh bt r tin, hoc nhp khu c nguyn liu r sn xut isoglucose m khng dng c cho ngi v gia sc. Cc tin trn gii thch ti sao M sn xut isoglucose pht trin mnh, m M mt nc phi nhp khu rt nhiu ng, ng thi sn xut 46% tng sn lng ng th gii. Cng nh Nht Bn, mt nc nhp khu ng ch yu v nhp ti 99% ng cho mnh. Cc nc ng u v mt s nc ang pht trin ngy cng quan tm n sn xut isoglucose (Argentina, Australia). ng Nam (Malaysia, Pakistan, Hn Quc) v chu M Latinh (Brazil, Chil, Mexico, Pru) ang xy dng cc x nghip v th trng m ra nhanh chng. Sn lng isoglucose nm 1988 M l 4,2 triu tn, Canada: 300.000 tn, Nht Bn: 800.000 tn, khi EEC: 1-2 triu tn, cc nc ang pht trin: 500.000 tn, cc nc ng u: 200.000 tn. Nu khng k EEC sn lng isoglucose 6 triu tn ca th gii nm 1988 s tng ng vi kh nng ti a ca th trng. Chc chn phng php mi s lm gim gi thnh sirofructose v tng tc thay th ng.Vic bn xi-r dng kh ngy cng tng s cnh tranh vi ng s dng trong ni ti.
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Chiu hng ny c nh hng quan trng trn phm vi th gii, c bit i vi nn kinh t v cn cn ngoi thng ca cc nc sn xut ng vn b thit thi do gi ng gim lin tc trn th trng th gii. 2.2. Cng ngh sn xut protein nh vi sinh vt C rt nhiu loi sn phm ch bin t protein nh tht hp, c hp, nc mm ngn ngy, rm bng, xc xch, lp xng, pat, cc mt hng thy sn lnh ng, qui m cng nghip. y chng ti ch gii thiu mt vi sn phm tiu biu lm v d. Sn xut protein n bo Vic nghin cu vi sinh vt qui m ln lm ra ngun protein cung cp cho con ngi v gia sc, gia cm c coi l mt bin php gii quyt nn khan him thc n c trong i chin th gii ln th nht. Ti Berlin, Delbruck cng cc ng s ca mnh ln u tin t chc nui cy ln nm men bia Saccharmyces cerevisiae. Sinh khi nm men ny c dng ch yu b sung vo xp v xc xch. Cc nm men thc phm (Candida arborea, Candida utilis) mt ln na gp phn quan trng vo vic b sung thc phm hng ngy ca ngi c trong i chin th gii ln th hai. (Theo Tp ch Scientific American, 1981). Hin nay, nm men c sn xut nh mt ph phm ca cng nghip bia, hoc c nui trn nh nhanh (Lactoserum) mt ph phm ca ngnh sn xut fromage. Sinh khi nm men cng dng lm thc n cho ln con cung cp loi acid amin khng thay th c l lysine. Trong nhng nm 1960, thut ng protein n bo (SCP: single cell protein) ln u tin c dng ch cc protein vi sinh vt sn xut bng cch nui cy ln nm men hoc vi khun c dng lm thc n cho ngi v gia sc, gia cm. C cht dng cho vic sn xut protein n bo l hydrocarbon du m, r ng. B thi ca cc nh my bt giy nui sng cc nm men Saccharomyces, Candida, Mycoprotein, (protein nm v d: Fusarium gramineum), cc loi to n bo: Chlorella, Scenedesmus, Spirulina, Sn xut protein t parafin du m Ma h nm 1913, gio s E.E. Uspenski trng i hc Tng hp Moskva tin hnh nghin cu cy tm ma (Urtica) v cy mc tc (Equiserum) mc trn than v trn nc. ng thy i khi trn thnh chu lm bng parafin, hay trng parafin xut hin khun ti nm mc v mt lp vng vi khun. Uspenski cy cc vi sinh vt ny v xc nh c lai lch ca n. l Aspergillus flavus. Cui nm 1922, hc tr ca ng l V.O.Tauson nghin cu iu kin pht trin ca nm trn parafin v sinh l hc ca qu trnh oxyha ny. Nm 1925, lun n tt nghip ca
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nh khoa hc tr tui ny S ng ha parafin bi vi sinh vt bo v thnh cng xut sc. Lc cng l lc xut hin cc thng bo ca Rahn, Tausz, Sohngen v Vaner v nhng trng hp oxyha parafin bi vi sinh vt m h quan st thy. Trc , nm 1695, Miyoshi pht hin c kh nng nh th nm, nhng hi cc cng trnh ca cc nh bc hc trn th gii hon ton khng xem xt ti vai tr ca vi sinh vt trong vic phn gii hydrocarbon trong t nhin cng nh nghin cu rt t v sinh l hc ca cc vi sinh vt ny. Tauson chng minh rng, parafin c th dng lm ngun dinh dng carbon duy nht cho mt nhm ln cc sinh vt. Cc nghin cu c c bit ch khi c s e da gay gt i vi nhn loi v nn i protein. Cc t chc nghin cu khoa hc ca Php, Anh, Italia, Nht Bn v M bt u y mnh vic nghin cu vn thu nhn protein t du m. Hng du m Bistish Petroleum (BP) ca Anh t kt qu nghin cu ca Champagnat ti La Viera (Php) quan tm n vic sn xut sinh khi vi sinh vt trn hydrocarbon t cui nhng nm 1950. D n ban u ngoi vic sn xut protein cn nhm mc ch kh sp cho du m. Du th La Veria cha t 10-15% parafin. Cht ny c nhng loi nm men Yarrowia (Candida) chuyn ha t cc ni ln men c b sung ammoniac (NH4+ + NH3) cung cp N2 v gi pH thch hp cho mi trng khng kh trng (Candida lipolytica ln u tin c tch t b thc vt Ha Lan nm 1928 c gi l Torula lipolytica sau i thnh Saccharomycopsis v n nay li i Yarrowia lipolytica. Nm men ny oxyha c hydrocarbon thnh acid bo nh enzyme hydroxylase. Cc acid bo s c phn hy tip thnh acetylCoA ri c chuyn thnh sinh khi nm men). Sau giai on sinh trng mnh m nm men c tch ra v phn du cn li c chuyn i, phn lipid chim n 10% ca sinh khi. u nm 1970, nhiu nhm ha hc v cng nghip du m, ch yu l chu u v Nht Bn quan tm n vic xy dng cc qui trnh cng ngh v xy dng cc nh my nhm sn xut ra protein n bo ca nm men hay ca vi khun t c cht l hydrocarbon, methan, hay methanol. Nh my c qui m ln u tin ca hng BP ra i Sarroch (vng Sardinia) vo cui nm 1975, sn xut nm men t n-parafin vi cng sut 100.000 tn/nm. Mt nh my khc ca BP Grangemouth (thuc Scotland) vi nhng ni ln men c khy trn cha mi trng c n-parafin thun khit. Sn lng protein n bo l 4.000 tn/nm. Cng ti Liquichimica Italia cng xy dng nh my sn xut t n-parafin ra
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protein nm men, acid amin, acid bo v acid citric (qui trnh ca Nht Bn) cng sut nm 1980 l 200.000 tn protein. Rumanie ti vng Curtea de Arges xy dng mt nh my sn xut protein n bo theo qui trnh ca cng ti Dainippon Ink. & Chemical (Tokyo). Nm men c dng l Candida parafinica cho pht trin trn 4 ni ln men hnh thp c sc kh dung tch 1.260 m3. C cht ln men l parafin. Khng kh sau khi c nn v lc s c a vo ni ln men sc kh v lm phn tn parafin. Sn lng 60.000 tn/nm (gi thnh 1000 USD/tn protein n bo vi tn thng phm Romiprot). Sn xut protein t methanol Sn lng kh t nhin to ln Bc Hi cng lm cng ti Shell International Petroleum quan tm n vic dng nguyn liu ny lm c cht cho vic sn xut protein. Kh thin nhin Bc Hi gn nh l kh methan tinh cht. Chng c dng tng hp ammonium trong ngnh ha du. phng s bc chy ca methan ngi ta oxyha thnh methanol nh phng php ha hc. Vic sn xut protein t methanol l phng php c s dng trong cc nh my ca cng ti Hocchst AG Cng ha Lin bang c v ca cng ti ICI (Imperial Chemical Industrics Lid.) Anh. Phng php ny cng c vin Nghin cu Khoa hc Kowait tip nhn v mt nh my sn xut th sinh khi nm men c trin khai nm 1982. Nhm cng ti ICI tham gia sn xut protein t nm 1968. Nm 1980, xy dng mt nh my Billigham (Anh) v cnh tranh vi u tng bng cch nng cao hiu sut chuyn ha methanol thnh protein. Chng vi khun c s dng Methylophilus methyloropus ASI c kh nng ng ha ammonium t dch nui cy bng cch chuyn ha n thnh acid glutamic nh enzyme glutaminsynthetase (GS) v enzyme glutamatesynthetase (COGAT). Vic ng ha ammonium bc xc tc nh GS cn c nng lng dng ATP. ATP c sinh ra khi oxyha methanol thnh CO2 v vic ny lm tiu ha qu nhiu carbon. mt s vi khun khc k c E. coli, c mt cch tng hp acid glutamic rt quen thuc i hi t nng lng hn v ph thuc vo mt enzyme khc glutamate dehydrogenase (GDH). Ngi ta tch c mt chng t bin ca vi khun ASI khng ng ha ammonium v thiu enzyme COGAT. Mc khc, cc gene m ha vic tng hp GDH ca E. coli c gn vo mt plasmid v chuyn vo t bo ca ASI. V vy, c th ng ha ammonium m chi ph nng lng thp hn. Cui nm 1983, cng ti ICI hp tc vi Lin X xy dng mt nh my sn xut protein cng sut 100.000 tn/nm v cn cung cp mt nm 200.000 tn methanol.
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Nhiu cng ti khc trn th gii qui m nh hn cng tin hnh sn xut protein bng phng php methanol ny. Sn xut protein bng cc c cht khc u nm 1960, trong vic sn xut protein t hydrocarbon cn c cc nh my ln men c xy dng sn xut sinh khi vi sinh vt t r ng v t b thi ca cc nh my giy. R ng ma hay r ng c ci l cht dch cha 50-60% phn t hydrate carbon nh b, c th ln men nh nm men Saccharomyces cerevisiae hay Candida/Torula utilis. Dch thi ca nh my bt giy cng cha mt nng thp cc ng c th ln men bng nm men. Nhiu nh my ln men c xy dng M v cc nc Bc u, dng dch thi ca bt giy sn xut protein, nh m gim vic lm nhim sng ngi bi dch thi ny. Loi nm men pht trin trn dch kim sulphite (Candida utilis) l loi nm men thc phm c dng lm cht iu v cho thc n v xp. chng nhim mi trng do sn xut bt giy v tng lng protein, nhiu cng ti Envirocon Lid., Vancouver, Cellulose Attisholz (Thy S), United Paper Mills Jamsankoski, cng ti Giy v Bt giy G.A. Serlachius Oy (Phn Lan) tin hnh sn xut protein dng trong chn nui. V sn xut protein t r ng th Cuba sn xut nm men gia sc (Candida utilis) t r ng ca nh my ng v t hn hp gia r ng v b ru. Nm 1986 c ti 11 nh my sn xut protein hot ng Cuba v 2 nh my khc sn xut men bnh m. By trong s cc nh my ny do cng ti SPEICHIM ca Php thit k, cc ni ln men vi qu trnh sc kh. Bn nh my cn li do cc cng ti o thit k. Tng cng sut hng nm l 130.000 tn, sn lng thc t l 90-100.000 tn sinh khi/nm. Sinh khi ny ch yu dng trong chn nui. Ngoi vic dng nm men ln men, Billigham (Anh) dng loi nm si Fusarium gramineum sn xut trong giai on u (1.000 tn protein/nm). Loi mycoprotein ny c cng ti Rank Hovis Mc. Drgall ca Anh bn ra th trng qua chi nhnh New Era Food (do cng ti ny v cng ti ICI lp ra). S dng mycoprotein c php dng vo thc phm cho ngi v protein chim 44% trng lng kh, saccharide: 19%, cellulose: 18% v lipid: 14%. T nm 1975, Azoulay gip cho hng Adour Entreprise (ty nam nc Php) phn lp mt chng nm men Candida tropicalis c th ln men trc tip sn m khng cn qu trnh thy phn ban u. C 2 kg bt Sn kh th c 1 kg sinh khi nm men (hay 7 kg sn ti c 1 kg sinh khi). Bt nm men sy kh >100oC phn hy acid hydrocyanic c trong sn (Manihot esculenta) chuyn thnh
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ammonium v acid formic. (cc loi sn khc nh M. aipi khng c hydrocyanic nu khng cn x l loi b c tnh ca chng). n , Thi Lan cng sn xut v xut khu sinh khi nm men t Sn. Ecuador v Colombia cng sn xut sinh khi nm men t chui km phm cht, khng xut khu c, bn hng nm 1 triu dollards M. Ch bin cht ngt t nhin Sn xut cc cht ngt t nhin bng nui cy m v t bo thc vt cng nh cc cht ngt khng phi ng bng phng php CNSH khc c th lm xu i tnh trng ca ng ma v c ci ng trn th trng quc t. Gi ng bt u h t nm 1985 do lng tiu th cc nc cng nghip gim nhanh, do cc nc xut khu sn xut qu nhiu v do cnh tranh ngy cng tng ca siroffructose (isoglucose) v cc cht ngt bn tng hp (aspartam) hoc cht ngt nhn to (acesulfam K). Theo ti liu ca Wolkstein nhan ng trc s tn cng tip ca fructose, polysaccharide v cc cht ngt khc xut bn nm 1986 ti M. Trong vng 15 nm, k t nm 1987 v trc, ti thiu c 8 cht gy v ngt ang c nghin cu v c tung ra th trng. Chnh ti liu kt lun: th trng M s t mc 154,3 t bng Anh (pao) cht ngt qui chun ra ng v vo nm 1990 vi s phn b nh sau: cc cht ngt c calo s gim t 129,8 xung 124 t pao qui ra ng v 109,6 t pao vo nm 2005. Cn cc cht ngt t calo hoc khng calo s tng t 17 n 30,5 t pao vo nm 1990 v 45,3 t pao vo nm 2005. D bo ny cho thy r t l quan trng ca cc cht ngt t calo (xem Nothias J.K., 1987). Cc cht ngt t calo gm nhiu cht c ngun gc khc nhau, t nhin hoc nhn to. Cht ngt t nhin c a chung hn v c gn nhn t nhin v v vy c dng nhiu hn trong thc phm. Th trng cha thc phm v nc ung t calo ang tng ln nhanh chng: 2,5 t USD doanh s nm 1982 v 41,2 ti USD nm 1990. Nhu cu tng nhanh ny li i km theo bi s ch nhiu hn ca ngi tiu dng n thnh phn thc phm. V vy cc cht ngt t nhin c a thch hn. Cn ghi nhn rng, cc cng ti thc phm, ha cht v ha dc c a quc gia ngy cng tham gia nhiu n s pht trin cc cht c v ngt mi nh Tate v Lyle (Anh), E.I. Du pont de Nemours, Monsanto, Pfiser (nm 1986 xin c giy php bn mt cht ngt tng hp mi tn l alitam M) v mt s cng ti ln Nht Bn.
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Tp ch M Bioprocessing Technology s thng 8 nm 1986 cng tng kt v cc cht ngt c ngun gc thc vt t nhin gm mt s loi cht khc nhau. Trong nm 1970, tp on Tate v Lyle ch bin ng v thc phm ca Anh xy dng Gana, Liberia, Malaysia cc n in trng cy Thaumatococcus danielli (h Narantaceae) tm trong rng m Ty v Nam Phi, c qu chm mu rt ngt, cha mt protein ngt l thaumatin. Hng ny sau nghin cu sn xut thaumatin bng phng php CNSH (Kenney M.J., et al., 1983). Gene m ha thaumatin c nhn dng trong vi khun E. coli bi cc nh nghin cu phng th nghim Unilever (Colworth, Anh) v cc nh khoa hc trng i hc Kent truyn vo cy thuc l. Gene m ha cho thaumatin cng c th truyn vo cc cy trng thc phm tng mi v sn phm. Cc nh khoa hc INGENE (International Genetic Engineering) Santa Monica, California nghin cu nhn dng v a gene ny vo t bo nm men (Saccharomyces cerevisiae). Bng cch , c th to thnh thaumatin nh nui nm men trn qui m ln, sau , phc ch phn t protein (phc hi 8 cu ni disulphite to iu kin cho protein c c cu trc v tnh cht t nhin ca n). Cng trnh ny c hng Beatrice US Food Corporation ti tr t nm 1982 mi nm 1 triu USD v ring nm 1986 l 1,8 triu USD; cng ti ny c c quyn bn sn phm lm ra trong khi cng ti INGENE d kin s thu c 25 triu USD tin bn bn quyn (theo Biofutur s 54 thng 2 nm 1987 trang 13). Thaumatin c ngt gp 2.000-2.500 ln ng saccharose (so cng trng lng) vi gi tr 16.500 USD/kg nm 1984. V ngt ca thaumatin ln chm v gi c lu. N cn c tc dng tng v mt s cht nh bc h cay. c im ny to thun li cho thaumatin c s dng lm ko cao su v trong cng nghip dc. Nm 1987, thaumatin c bn o, Thy s, Nht Bn v Anh, trong khi M n ang c xem xt c tnh trc khi cho php bn. Cc nc khc cng ang trong qu trnh cho php bun bn thaumatin nh l cht ngt. Monellin l mt protein ngt khc chit tch t tri cy leo Doscoreophyllum cumminsii (h Menispenneaceae) tm thy vng nhit i m Ty v Trung Phi. Thaumatin v monellin l 2 loi protein ngt c bit, ngt gp 100.000 ln ng nu tnh theo phn t lng v gp vi nghn ln nu tnh theo trng lng. Ngi ta chng minh rng, hnh th nguyn gc l rt quan trng i vi v ngt mc d c hai loi protein u rt ngt, song gia chng khng nhng ging nhau ng k qua thng k trnh t acid amin ca chng. Mc d khng ging nhau v trnh t nhng cc khng th ca thaumatin vn ginh git monellin, song
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li khng ginh git loi monellin khng ngt b ci bin ha hc v ngc li (xem Ogata C. et al., 1987). tm hiu c s cu trc ca cc quan st trn, Ogata v ctv khoa Ha trng i hc Tng hp California, Berkeley xc nh cu trc ca monellin phn gii 3Ao, sau khi thng bo cu trc tinh th ca thaumatin nm 1985 (Ogata et al., 1987). Monellin gm 2 chui peptid, chui A gm 44 gc v chui B gm 50 gc. Khung cu trc tng th ca monellin v thaumatin khng c g ging nhau ng k c. Cc protein ny ch c 2 khu vc nh (c hai u l tripeptid) nm trong vng nt m, l ging nhau v trnh t acid amin. Cc nghin cu sinh ha v min dch cng c tin hnh nhm xc nh cc khu vc c th l chu trch nhim v phn ng khng th giao cho v lin kt th cm v ngt. Ch bin c tht hp, xc xch, gi, lp xng, nem chua Tht c cha cc hp kn v thanh trng nhit cao (115o 121 C khong 40-69 pht ty tng loi tht v s lng tp khun trn tht) Tht ng hp phi l loi tht tt t gia sc, gia cm khe, tht cn ti, cha c du hiu h hng. Trc khi thanh trng cn phi kim tra vi sinh vt c trong tht ttheo 2 tiu chun: tng s vi khun v bo t cc vi khun hiu kh a nhit. Trong qu trnh thanh trng, hu nh cc vi sinh vt gy bnh cht hon ton. Cc vi sinh vt ph hy tht cht phn ln. Cc bo t ca vi khun Bac. subtilis, Bac. mesentericus, Cl. botulinum c th vn cn sng. Trong hp tht, bo t Cl. botulinum c th pht trin sinh ra c t gy ng c tht. Trong tht c nhiu m, t bo vi sinh vt trong mi trng m bn vi nhit. Cho nn, ch thanh trng cn phi ch . Xc xch, gi, lp xng. cc sn phm ny l thc phm n trc tip khng cn qua un nu li. V vy, yu cu v sinh nguyn liu tht phi cao, tht loi tt, ti. Cng cn ch n cht lng nguyn liu ph nh bt, mui n, dim tiu (KNO3), gia v. Tht loi b gn, xng, cho vo xay mn ri trn mui nhit thp trong khong thi gian ty theo phng php ch bin. Ngoi mui cn cho thm dim tiu gi mu tht, thm ng cho mm tht, thm gia v (lm lp xng th thm hng lu). Sn phm c luc chn. Bo qun thng hoc bo qun lnh. Sn phm c th sy kh hoc xng khi. Trong vi khun c nhng chng lm hon thin mu sc cho xc xch. Mt s nc dng vi khun dng thanh khit cho vo xc xch: Pediococcus cerevisiae, Micrococcus T.53, L. platarum, M. aquatilis, M. auranticus, Str. Lactis, Micrococcus spp. Xc xch, lp xng, gim bng, gi, ch,c mt lng nc
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khng ln (30-40%), c v bc, c cc cht c ch vi khun (mui, dim tiu, gia v, cc cht trong khi) nn c th bo qun c mt thi gian. Nem chua. Ty theo tp qun ca mi nc, mi vng m cch lm c khc nhau. V d tm chua Longanisa ca Philippine bao gm tht nc 70%, m thi nh 30%, mui 2% so vi tng s tht, ng tinh 2%, gim 2%, ru vang 2%, ht tiu en 0,6%, ti gi nh 0,6%, potassium nitrate 0,05%, hn hp phosphate 0,15%. Vi khun ln men lactic t nhin c mt trong nguyn liu ban u. p 2-3 ngy trong t lnh, ly ra ngui cho ln men nhit 30-35oC trong 12-24 gi, em dng. Ln men tm, c v cc loi thy sn khc C tm mui C , tm mui l sn phm kh ph bin. Phng php mui tm, c bo qun c dng t thi c xa. Dung dch mui c kh nng to thnh p sut thm thu cao. Dung dch mui 10-20% c p sut thm thu 200 atm. T bo nguyn liu cng nh vi sinh vt b co nguyn sinh lm cho vi sinh vt hoc ngng hot ng. Cc qu trnh bin i cc cht khng thc hin c chc nng bnh thng. V vy mt s vi sinh vt gy hi khng hot ng c. Trong qu trnh mui tm, c c th xy ra nhng bin i ha sinh lin quan n tc dng ca protease v oxyha- kh cng nh s hot ng ca vi sinh vt c trong tm c. Tht ca mt s c mui tr nn mm, ti ngon v c mi v thm du. V vy, tm c mui chn v c th n trc tip khng cn phi qua nu nng. Gn y mt s nc bo qun c dng ng bnh. Nguyn liu em xay nh v bp mui. Sau em p ri phi hoc sy kh. Sn phm g c di ngy c trong iu kin kh hu nhit i. Trc khi nu, em ngm vo nc nng gim mn. Cc nc ng Nam chu dng tm, c mui kh ph bin nh tm mui thnh Balao Balao (Philippine), tm mui Kung Chom (Thi Lan), Tm mui Saeoojeot (Triu Tin), mm c thu Burong (Philippines), trai mui hoi Malangpu Dong (Thi Lan), tm chua Hu, mm nm, c mm cc loi, nc mm (sn xut khp c nc trong c nhng vng lm nc mm truyn thng tr thnh nhng mn hng c sn nh nc mm Phan Thit, nc mm Ph Quc, v.v.). 2.3. Sn phm t rau qu gim bt s tht thot cc sn phm rau qu, hin nay ngi ta cng ng dng nhiu hnh thc ch bin khc nhau nh sn xut cc loi rau qu hp, sy kh qu qui m ln v hin i, lnh ng rau qu, mui, ln men rau qu ri ng hp. Sau y l mt vi v d v vic ch bin rau qu
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Bo qun v ch bin rau qu bng phng php lnh v lnh ng Phng php lnh ng rau qu c bo qun 0oC s km hm qu trnh sinh l, sinh ha v hot ng ca vi sinh vt, ng thi lm tng hng v cho rau qu. Thng thng ngi ta bo qun rau qu trong kho lnh t 2 n 2oC. Cng ty theo phm cht ca sn phm m nhit bo qun c khc nhau. Ty theo mc x l nguyn liu c cch phi ch, rau qu lnh ng c cc dng sau: + Rau t nhin lnh ng + Rau rt nc mui lnh ng +Qu t nhin lnh ng + Qu trn nc ng lnh ng +Nc rau qu lnh ng +Rau qu nghin lnh ng. Sau y l mt vi qui trnh lnh ng rau qu.
Qui trnh lnh ng chui
Chui gi 95-98% Gim chn Ct ri tng tri Ngm, ra, st trng Ct khc, ct u Bc v ro nc
Ngm tm chng oxyha ro dung dch ngm tm Cn, cho vo ti Lm lnh ng -35-40oC ng kin Bo qun 18oC
Qui trnh lnh ng c chua

C chua chn 60-80% Chn, ra, st trng Rt cung Cn, vo t, dn ti Chy, ro nc Ra li bng thuc st trng Lm lnh ng nhanh -35 n 40oC ng kin Bo qun -18oC
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Qui trnh lnh ng hn hp rau

BP CI SP L Chn Chn Ra,st trng Ra,st trng Thi si Ct thnh mnh nh Chn Chn C RT HNH Chn Chn Ra,st trng Bc v ngoi Co sch v Ra Ra sch Thi si nh ngang To hnh (lt trn bu dc, si, lt hoa, ht u) Chn Chn
Phi trn cc cng thc khc nhau (k c thm gia v) Cn ng ti, dn ming ti Lm lnh ng nhanh -15 n 40oC ng kin Bo qun -18oC
2.4. Ch bin thc phm chuyn gene t hng nghn nm qua, loi ngi bin i di truyn cy trng nh gi ging tt t v trc qua v sau; cy c chua di thnh cy c chua hin nay; cy ng di bp trn di 2,5 cm thnh qu c bao di 30 cm. Hng trm ging mi c to ra bng chiu x hoc dng ha cht gy t bin. Mt trong nhng bin php hin nay l k thut nui cy m v t bo, to phi soma, ht nhn to, to dng soma trong nui cy m v t bo, protoplast, c bit s dng cng ngh gene trong ti t hp DNA.
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C nhiu ging cy mi c to ra theo mun ca con ngi, nh to ra cy va n qu, va n c (qu c chua v c c rt hoc c cua khoai ty trong cng mt cy, mt chm qu nho gia c qu c chua xen vo m qu c chua ny li t thi hng, v.v.), cy ng chuyn gene chng su hi v.v..
Ng chuyn gene khng su hi
Cy khoai-c (Pomato)
Hnh VII. Cy trng thc phm chuyn gene Hoc trong ht nhn to, ngi ta a vo v ht cc loi vi khun c nh N2, khi cy trng thnh, vi khun ny s ly N2 t khng kh cung cp phn m cho cy . Cng bng cch tng t, ngi ta a mt s thuc tr su hoc tr c di vo v ht nhn to bo v cho cy khi b su v c di ph hoi. Ht nhn to cng d dng np cc gene l vo nhm to ra cc ging mi c c tnh mong mun. V chuyn np gene, trn th gii cng c nhiu thnh tu. Ring Vit Nam, vin Cng ngh Sinh hc, vin Sinh hc Nhit i thnh ph H Ch Minh thuc vin Khoa hc v Cng ngh Vit Nam, vin Di truyn Nng nghip TW, vin La ng bng sng Cu Long thuc b NN&PTNT chuyn np gene chng chu ry nu vo cy la bng sng bn gene v c phng php gin tip bng plasmid Agrobacter tumefaciens cng t kt qu. Nhiu cng trnh nghin cu chng chu su bnh, bo v cy trng nh dng vi khun Bacillus thuringiensis (Bt) l loi c kh nng sn sinh ra protein chng cn trng. Ngi ta cng a gene vo to enzyme lm thoi ha thnh phn tin cht hnh thnh ethylen, nh vy s lm chm s chn v h hng. (Cng trnh ca Kishore v Harry Klee
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Monsanto). Cc nh khoa hc M cy vo cy mt gene vi khun sn sinh ra mt cht chitinase tiu dit t bo nm (cy vo c chua, khoai ty, rau dip v cc ging cy tng t nhng cha lm c vi la, la m, ng v cc cy c ht khc). Bn cnh vic nng cao nng sut cy trng, ngi ta cng nng cao phm cht ca chng bng cch a gene to enzyme mi nh G. Glili dng vi khun E. coli chit xut ly gene m ha loi enzyme tng hp lysine, sau a vo khoai ty v thuc l lm cho hm lng lysine trong khoai ty c tng 5 ln, l tng 4 ln v thn tng 3 ln. Ngi ta cng a 9 acid amin khc vo khoai ty lm cho cht lng khoai ty tr nn qu gi. Gia thp k 90 ca th k trc, Brazil, cng ti Sinh hc a d n ghp mt gene ca lc cho u tng nhm lm giu m cho u tng, b b v u tng ny gy d ng cho ngi. Nh vy, gia nhng nm 1990, cng ngh gene bt u a vo hn 50 sn phm mi c ng dng 13 nc vi din tch 52,6 triu ha nm 2001, tng 50 ln so vi nm 1996. Trong nhiu nht l u tng 34,9 triu ha, bng 64% din tch u tng th gii, ng 6,1 triu ha v 3,3 triu ha cy ci du. Ngi ta ra c hn 10 ging cy trng mang gene mi. n nm 2004 c n 81 triu ha trng cy chuyn gene 17 quc gia v d tnh vo nm 2010 din tch trng cy chuyn gene s tng ln n 150 triu ha vi khong 15 triu ngi trng ti 30 nc trn th gii. V vt nui cng c nhiu thnh tu. V d nh tim DNA vo trng chuyn i gene MT-hGH pht trin thnh blastocyste, cy na non bo vo gia sc to ra nhng con sinh i, dng hormone tng trng ti t hp gene ca b tng sinh l sa ( M lm tng 15% hm lng sa trong b), to b c v b ci theo mun, v.v. Bng k thut ln men, Elizabeth Stiton, Danny Lactaire a gene sn sinh vi khun ln men vo h di truyn con b sa, chuyn gene di truyn ca b sa thnh b cho yaourt. Vn ang tranh ci hin nay l sn phm chuyn gene c c an ton thc phm hay khng? c nhiu tranh lun v vn GMO (Geneticaly midified organisms c ngha l c gene ci bin hay chuyn gene). C ngi cho rng, sn phm chuyn gene hin nay nh gene ng BT, u tng BT, du ci BT l nhng sn phm k diu, kt qu ca cng ngh gene. iu ny rt ng hoan nghnh. Hng triu nhn dn M ang n loi sn phm ny, nhng chu u v nhiu nc khc li ty chay. y l mt nghch l khng th gii thch c C ngi cho rng, nhng nc phn i u c l do ca h. Nu nh ng vi kin phn i nh vy, ai m bo l sn phm ny khng c hi cho sc khe
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c con ngi? C ngi li ni: cng ti Cng ngh Sinh hc c tm c nht ca th gii Monsanto nu ln tnh trng bng n dn s trn th gii trong th k ti, i hi din tch nng nghip hin nay l 1,5 t ha phi tng ln 2,8 t ha. Nhng cha phi l bin php hu hiu gii quyt vn lng thc cho loi ngi. Ch c da vo mt nn nng nghip p dng cc ging chuyn gene mi c th nui sng c 10-12 t ngi, li tiu hao t nng lng v chng c nhim mi trng. Ngi khc li ni: hin nay M, cc sn phm chuyn gene hu ht do cng ti t nhn sn xut ra, h cp tin cho cc vin nghin cu, cc trng i hc nghin cu ti cho h. Cc cng ti u chy theo li nhun. H tuyn truyn rm beng, chc g tt c sn phm u phc v li ch lu di cho con ngi? Li kin tr li: hy c ni n li ch cho nh nng. Cc sn phm c gene vi sinh vt BT c th chng c su bnh, nh nng khng phi dng nhiu thuc ha hc phng tr, tit kim c chi ph, chng nhim sn phm, chng nhim t, nc v khng kh. kin chng li: nhng chnh gene li to ra c hi cho sn phm th sao? c g chng minh l v lu v di bin i an ton cho ngi? Mi y tu ch u tng t M b dn Anh ty chay y v.v. v v.v. ng l cn qu nhiu tranh ci. V chuyn m thc thi chuyn gene, trong Tp ch Th gii mi, Thiu Lm vit bi Thc phm chuyn gene - Th trng khng ai kim sot trong trch dn u tin vit: Sau khi n mn bnh bt ng nhn tht g, b Grace Booth 35 tui California (M) cm thy nng trong ngi, bun nn, sau b tiu chy, sng v mi, kh th. Cc bc s chn on b b sc phn v nn tim thuc khng d ng, truyn dch v cho ung thuc khng histamine Benadryl. Nm gi sau, b Grace Booth xut vin. Sau ny b mi bit b b d ng vi ng chuyn gene StarLink dng lm nguyn liu ch bin bnh bt ng nn gi n n kin c quan Qun l Thc phm v Dc phm (FDA). T nhiu nm nay, thc phm chuyn gene b nghi ng c cha nhiu protein l c th gy hi sc khe cho con ngi. Tuy nhin, y l ln u tin ngi tiu dng i kin. Thng 3 nm 2001, tp ch Discover ca M a ln trang ba v ng StarLink gy d ng. FDA cng chun b xt nghim mu mu ca 44 ngi lm n kin ca Aventis nh sn xut Ng chuyn gene StarLink. Tc gi Thiu Lm vit trong tp ch Th gii mi (2001, trang 2125) nu nhng mc chnh nh sau: Bnh bt ng gy d ng. Tc gi a ra nhiu trng hp M, ngi dng bnh ng chuyn gene b d ng. Ng StarLink bt u xut hin trn th trng M vo nm 1998. Ging ng ny mang gene vi khun Bacillus thurigiensis c kh nng tit
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c t dit bm ng gip cy Ng khng b ph hi. Tuy nhin, trong ng StarLink c protein Cry9C, mt loi protein khng b cc enzyme c bit trong b my tiu ha con ngi phn hy. Trong s 44 trng hp kin co v ng StarLink, trung tm Kim sot Dch bnh M (CDC) ch n 26 trng hp v a ra kt lun ban u: 11 trng hp xy ra triu chng d ng v 7 trng hp trong s ny chc chn c lin quan trc tip n vic n thc n bng bt ng chuyn gene StarLink mang nhn hiu Taco Bell hoc Mission Foods. Tuy nhin CDC vn cha khng nh Cry9C c trong thc n hay khng v FDA s tip tc lm cng vic ny. Mc chnh khc Thiu Lm vit: Khng ai dm chc thc phm chuyn gene gy ng c v ngc li. Thc phm khng chuyn gene: kh phn bit tht gi. Ngi ta biu tnh chng cy chuyn gene Brazil. ng rung trng cy ci du chuyn gene b ph b Php. ng, thc phm chuyn gene l vn cn ang tranh ci.Thi gian qua, cc nc EU khng chu nhp thc phm chuyn gene ca M. Sau ny h yu cu thc phm chuyn gene phi c l lch chuyn gene. (Gn y c thng tin cho rng EU quyt nh cho nhp thc phm chuyn gene tr li nhng khng gii thch l do). C mt s ngi yu cu gn nhn hiu cho sn phm mi c cy ghp gene. Nhng sn phm mi khng khc my v c bn ca sn phm c truyn. V li, t gia nhng nm 1990, hn 60% sn phm lng thc, thc phm ch bin cc siu th Hoa K gm c pizza, khoai ty lt, xi-r ng, bt m, u tng, ci du, u l sn phm cng ngh gene mi. Do gn nhn hiu l khng cn thit. Sn phm c gene mi c hi cho mi trng khng? Dng lan truyn gene thng qua ht phn, ht ging t ging ny qua ging khc l mi lo ca mt s nh khoa hc. Vic lan truyn t cy trng sang c qua gi, ong bm hoc cc yu t khc l c th xy ra. Chng ti ngh, l lch gene l quan trng. Khng phi gene no cng gy phng hi cho con ngi. V d nh trn ni, tng thm mt s gene to enzyme, t to ra sn phm c nhiu acid amin khng thay th lm cho cht lng ca sn phm tng ln nh tng 9 acid amin cho c khoai ty, hoc chuyn thm gene sn sinh vi khun ln men vo h di truyn ca b sa, chuyn b sa thnh b cho yaourt v.v. th u c hi! Cn vic a gene chng su bnh vo t bo th r rng c hai mt: cy s khng b bnh v cho nng sut cao hn; sn phm to thnh c nhiu hn. Ngay c khi b bnh, bn thn c th mun chng li iu kin bt li , sinh vt phi tit ra cc khng th c dit tr nhng tc hi gy ra. Cc khng th vn cn lu li trong c th, nu nng cao
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cng c hi cho con ngi. Trong trng hp con ngi can thip vo bng phng php ha hc hoc phng php chuyn gene chng su bnh, nu nh nng cao l c hi cho sc khe con ngi. Vn l nng s dng v s tch ly cht c hi c th. Nu nng cao th phng php phi loi tr, nu nng thp cho php th nn s dng. V vy cn phi tin hnh phn tch v phi c l lch ca i tng cn tiu dng. Gn y, trong Tp ch Th gii mi s 595 ra ngy 26/7/2004 trong bi Kh nng k diu ca chp sinh hc, tc gi Ngc Nhn vit:cng ti Apibio ca Php bc u a vo th nghim mt loi chp sinh hc c kh nng pht hin nhanh cc loi thc phm bin i gene. Ngi ta s dng chp c gn DNA c trng ca loi khng bin i gene Khi th nghim, thit b s so snh gene ca loi thc phm nhp khu vi gene mu trong chp. Nu hai nhnh DNA nhn ra nhau, iu c ngha l thc phm nhp khu l th thit. Tim nng sinh hc mi rt to ln, nhng n cng khng th mang li cc ri ro. Ph nhn hay ch da vo k thut mi u l sai lm. Chng ta cn phi phn tch cn k v tr li. u, nh th no, ti sao chng ta li s dng cc sn phm sinh hc mi v nu chng ta th nghm rng ri, phn xt khn ngoan, cn nhc li hi, l nhng iu cn thit hn c cho nhng ai cn n n. (trong mc ny chng ti c s dng thm ti liu Sn phm bin i gene ca T T trong bo Nhn dn s 65/92002. TI LIU THAM KHO i Duy Ban, L Th Cm Vn, 1994. Cng ngh gene v cng ngh sinh hc ng dng trong nng nghip hin i. Nxb Nng nghip H Ni. L Don Din, 1994. Cng ngh sau thu hoch trong nng nghip Vit Nam: Thc trng v trin vng. Nxb Nng nghip H Ni. Nguyn Ln Dng, 1992. Tm hiu v cng ngh sinh hc, Nxb Gio dc H Ni. Trng Vn Lung, 1995. Chuyn Cng ngh sinh hc. T sch i hc Khoa hc Hu Trng Vn Lung, 2002. Cng ngh sau thu hoch. (H hp vi vic ch bin v bo qun thc phm). T sch i hc Khoa hc Hu. Ngc Nhn, 2004. Kh nng k diu ca chp sinh hc, Tp ch Th gii mi, Nxb Gio dc H Ni, s 595, tr: 52-55.
1. 2. 3. 4. 5. 6.
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7. Phan C Nhn, Trn nh Min, 1997. Tm hiu cng ngh sinh hc

hin i. Nxb Gio dc, H Ni 8. Nguyn Hu Phc, 1998. Cc phng php ln men thc phm truyn thng Vit Nam v cc nc trong vng. Nxb Nng nghip thnh ph H Ch Minh. 9. H Sng (ch bin), 1982. Vi sinh vt trong bo qun v ch bin thc phm. Nxb Nng nghip H Ni. 10. L Ngc T (ch bin), 2000. Ha sinh cng nghip, Nxb Khoa hc v K thut H Ni 11. Trn Cm Vn, Bch Phng Lan, 1995. Cng ngh vi sinh v bo v mi trung. Nxb Khoa hc & K thut. Trung tm Giao lu quc t v Vn ha Gio dc v Khoa hc (CCES), H Ni. 12. Albert Sasson,1988. Biotechnologies and development Cng ngh sinh hc v pht trin. Ngi dch: Nguyn Hu Thc, Nguyn Ln Dng v mt s dch gi khc. Nxb Khoa hc & K thut H Ni. 13. Bezborodov A.M., Moxolov V.V., Rabinovitch M.I., Nguyn Vn Uyn, Ng K Sng v nnk, 1994. Cng ngh sinh hc v mt s ng dng ti Vit Nam, Tp I, II. Nxb Nng nghip H Ni. 14. Clive James, 2004. Tm tt bo co s 32 ca ISAAA v tnh trng cy trng chuyn gene/cng ngh sinh hc c trng v mua bn trn th trng th gii trong nm 2004.Bn tin Cng ngh Sinh hc thng 01 nm 2005. www.biotechvn.com,vn
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4
CNSH VI VIC BO V MI TRNG Chng VIII
Cc hot ng sn xut nng lm ng nghip v cng nghip cng nh thc trng v nhim mi trng
Khoa hc k thut ngy cng pht trin khin cho con ngi vn xa hn, rng hn v c m chinh phc c tri t. Dn s tng nhanh, lng thc thc phm ngy cng khan him khin cho con ngi tm mi cch khai thc cn kit ngun ti nguyn thin nhin. Con ngi bt chp tt c, th ri chnh con ngi ang phi gnh ly hu qu ny. l nn hn hn, l lt, bnh tt, nhim mi trng,... Bc sang th k XXI l th k ca cng ngh sinh hc v cng ngh thng tin, ng trc tnh hnh mi trng ang ku cu th ngnh cng ngh sinh hc l mt trong nhng ngnh i u trong cng tc bo v mi trng. CNSH ang ng mt vai tr ht sc to ln, thc y cc ngnh cng ngh khc cng pht trin. Mt trong nhng vn kh khn hin nay l nn nhim mi trng sng, n c xem nh l c bn dn n cc thm ho v mi trng.
1.Mt s nh ngha
1.1.nh ngha mi trng Theo nh ngha ca UNESCO (1981) th mi trng ca con ngi bao gm ton b cc h thng t nhin v cc h thng do con ngi to ra, nhng ci hu hnh ( th, h cha...) v nhng ci v hnh (tp qun, ngh thut...), trong con ngi sng bng lao ng ca mnh, h khai thc cc ti nguyn thin nhin v nhn to nhm tha mn nhng nhu cu ca mnh. Nh vy, mi trng sng i vi con ngi khng ch l ni tn ti, sinh trng v pht trin cho mt thc th sinh vt l con ngi m cn l khung cnh ca mt cuc sng, ca lao ng v s ngh ngi ca con ngi.
1.2 nh ngha nhim mi trng nhim mi trng l s nhim bn ca mi trng lm cho mi trng khng cn trong lnh, sch s. S nhim bn c th xy ra mi trng t, mi trng nc, mi trng khng kh do cc tc nhn gy nhim tng ng vi tng loi mi trng trn. 1.3 nh ngha cht thi Cht thi bao gm cht thi rn, lng, kh c thi ra sau qu trnh s dng ca con ngi v b thay i tnh cht ban u ca chng. Thng thng cht thi c phn loi theo ngun gc pht sinh ra chng. l c s cho vic la chn cc bin php hoc cng ngh x l. Cht thi c phn thnh cc loi: + Cht thi sinh hot: cht thi t cc khu dn c, khu vc hot ng thng mi, cng s, trng hc... +Cht thi cng nghip: cht thi t cc nh my ang hot ng, c c cht thi sinh hot nhng trong cht thi cng nghip l ch yu. +Cht thi th: l nhng cht thi trong cc h thng cng thot ca mt thnh ph. Nguyn nhn gy nhim mi trng: C nhiu nguyn nhn gy nhim mi trng. C nguyn nhn chnh do hot ng ca cc qu trnh t nhin din ra trong mi trng nhng cng c nguyn nhn li xut pht t nhng hot ng ca con ngi gy ra. * S pht trin ca cc ngnh cng nghip: Hu ht cc nh my u thi ra cc loi kh c lm nhim bu kh quyn. Lng kh CO2 do cc nh my thi ra lm nhim nghim trng mi trng kh, gy nn hiu ng nh knh. Nhng c bit nghim trng l CFC, c s dng nhiu trong cng nghip lnh, l tc nhn ph v tng ozone bao quanh tri t. Cc hot ng cng nghip cn thi ra mi trng mt lng nc thi cng nghip, l nguyn nhn quan trng gy nhim ngun nc v nhim c t.Trong cng nghip dc phm, cht thi t hot ng sn xut cht khng sinh thng cha ng t bo vi sinh vt, mt vi sn phm bin thi cng nh thnh phn khng dng n ca mi trng nui cy cng gp phn ng k vo vic lm nhim mi trng nc. Cc nh my cn thi ra cc cht rn, cc ph liu ca cng nghip cng l ngun gy nhim quan trng. * Hot ng nng nghip: Vic lm dng thuc tr su, phn ha hc l mt trong nhng nguyn nhn gy nhim mi trng quan trng. Thuc tr su, phn ha
hc lm nhim c mi trng t, mi trng nc ln mi trng khng kh. Thuc tr su l nhng ha cht c nn khi s dng nhiu n khng c phn hy ht m cn tn ng trong cc sn phm nng nghip. Khi con ngi, gia sc s dng nhng sn phm ny th c th mc phi nhng bnh nguy him nh ung th, bnh di truyn, thm ch rt d b ng c thc phm. Ngoi ra thuc tr su cn tiu dit cc sinh vt c li khc nh chim, ch, cn trng c ch... lm mt cn bng sinh thi, lm suy gim a dng sinh hc. Vic s dng phn bn ha hc nhiu, ko di t nm ny qua nm khc gy ra nhng tc hi nghim trng cho mi trng sinh thi: lm xu tnh cht ha l ca t, lm xu tnh cht vt l ca t, lm cho t b chua hay b kim ha, tch ly trong t nhiu ion c do cy khng s dng hay khng s dng ht. T nhng nguyn nhn trn lm cho t ngy cng xu i, cn bng sinh thi trn ng rung b ph v. Ngoi ra, c phn ha hc cung cp cho nng nghip cc nh my sn xut phn bn trn th gii hng ngy thi ra khng kh, nc, t nhng cht rt c hi lm nhim mi trng. * Ph rng: C th xem ph rng l nguyn nhn c bn, l ngun gc ca nhiu nguyn nhn khc lm mt cn bng sinh thi v nhim mi trng sng. Rng chnh l ci my iu ha t l kh O2/CO2 khng l trn tri t gip cho vic duy tr s sng trn tri t. Rng chnh l l phi ca tri t. Rng cn to nn mt thm thc vt gi nc khng l ngn chn xi mn, l lt...Rng cn l ni bo tn tnh a dng sinh hc, bo m s cn bng trn tri t. Th nhng hin nay rng ang b tn ph nghim trng, din tch rng ngy cng b thu hp do hot ng ca con ngi. Hng nm c khong 10 triu ha rng b con ngi trn th gii ph hy vi cc mc ch khc nhau. Ph rng gy ra nhng hu qu nghim trng. - Nn ph rng, t rng l nng ry lm tng CO2 gy mt cn bng t l O2/CO2 c nh hng trc tip n s sng trn tri t. - Rng u ngun b ph lm gim kh nng gi nc l nguyn nhn chnh gy ra l lt, xi mn. - Rng b cht ph lm mt thm thc vt bao ph tr thnh t trng i trc lm cho s xi mn xy ra nghim trng. Cht dinh dng trong t b tri theo dng nc, t cn dn cht dinh dng.
- Rng b ph lm gim tnh a dng sinh hc. S suy gim tnh a dng sinh hc trn tri t dn n s dit vong ca nhiu loi ng thc vt t lm cho cn bng sinh thi b ph v. * Hot ng ca con ngi: X hi cng vn minh, dn s cng tng th ngy cng xut hin nhiu yu t gy nhim mi trng. - S bng n dn s ko theo qu trnh th ha ngy cng din ra mnh m m mi trng th l mi trng b nhim nng. - Sinh hot ca con ngi to nn lng rc thi, nc thi ln gy nhim mi trng. Dn s cng tng th nguy c ngy cng ln. - X hi cng vn minh, con ngi cng c nhu cu s dng nhiu loi phng tin gy nhim mi trng nh xe c ng c, t lnh... nhim do xe c thi ra trong mi trng khng kh ngoi CO 2 ra cn c c Pb. * Nhng nguyn nhn khc: Bn cnh nhng nguyn nhn thng xuyn trn cn c nhng nguyn nhn khng thng xuyn nhng khng km phn quan trng cng gp phn gy nhim mi trng - Nn chy rng t nhin. - Nn m tu ch du. - Cc v th v kh ht nhn... - Chin tranh ha hc, vi trng... - Tai nn cc nh my ht nhn. Nm bt c nhng nguyn nhn gy nhim mi trng, con ngi tm ra nhng gii php thch hp ngn nga s nhim mi trng. Gii php u tin v cng c ngha quyt nh l ngn chn ngay nhng nguyn nhn do ch quan con ngi to ra. ng thi xy dng nhng gii php khoa hc ngn chn v x l hu qu ca s nhim mi trng. Trong nhng gii php khoa hc th cng ngh sinh hc ng gp vai tr ch o, quyt nh nht.
2. S nhim mi trng
* nhim mi trng khng kh: Theo TCVN 5966-1995 s nhin khng kh c qui nh nh sau: l s c mt ca cc cht trong kh quyn sinh ra t hot ng ca con ngi hoc t cc qu trnh t nhin v nu nng ln v thi gian lu chng s nh hng n s thoi mi d chu, sc khe hoc li ch ca ngi hoc mi trng CO2 thi vo kh quyn ngy cng tng cao : - Nm 1950 : lng CO2 thi vo khng kh l 12 triu tn/nm - Nm 1980 : lng CO2 thi vo khng kh l 150 triu tn/nm - Nm 1990-1991 : lng CO2 thi vo khng kh l 3000 triu tn/nm
- Nm 1996 : lng CO2 thi vo khng kh l 5200 triu tn/nm - Nm 1998 : lng CO2 thi vo khng kh l 6000 triu tn/nm Tuy nhin, hi ngh thng nh v mi trng hp ti Rio De Janeirio vo thng 6/1992 yu cu gim thiu mc thi CO2 trong nhng nm cui thp nin 90 ch bng mc nm 1991 l 3000 triu tn/nm. Ngun gy nhim khng kh C khng di 100 cc tc nhn gy nhim khng kh. Tuy nhin c 9 nhm chnh gy nhim khng kh bn ngo Carbon oxides: CO v CO2 Sulfur oxides: SO2 v SO3 Nitrogen oxides: NO, NO2 v N2O Cc hp cht d bay hi (VOCS): c hn 100 hp cht khc nhau nh: CH4, C6H6, CFCS. Cc cht l lng c bit (SPM): c hn 1000 dng khc nhau nh bi rn, cc dng git lng, du, H2SO4, Ch, Asen, Cadimium, PCBs, DDT v cc loi thuc tr su... Cc cht quang ha nh: PANS, NOX v cc hp cht hydrocarbon d bay hi di tc dng ca nh sng mt tri. Cc cht phng x: Radium 222, Iode 131, Strontium 90, Phetonium 239 v cc ng v phng x khc c trong kh quyn. Nhit: nhim nhit xut hin khi nng lng b bin i t dng ny sang dng khc nh khi s dng cc cht t l nguyn liu ha thch, cc nh my, cc khu cng nghip v cc cht t bng g ci. Ting n: Ting n nhim bi cc ng c, xe c, my bay, tu ha, cng nghip ch to, cng nghip xy dng, my xn c, my ht bi, radio, cassette v cc hat ng khc. Cc nh my ha cht, cc hot ng c tnh cng nghip thi vo mi trng nhng cht c hi nh dioxydsulfur , CFC (Clore Flore Carbon) lm bo mng tng ozone, tng hiu ng nh knh Cng nghip to ra nhiu ngun nhim khc nhau. V d: sn xut giy gy ra bi v hn hp hydrogen, ch bin ht iu to ra bi H2S dng hi v H2SiF6. Nh my lc du to ra bi, mi hi v phenol. Nh my thuc l to ra bi, mi hi nicotine, cc nh my hp cht SO2, NO2, CO2. Nh my supper phosphate to ra HC, SO2, CO2, NO2. Thnh ph H Ch Minh c gn 1000 nh my cng nghip, 30.000 c s sn xut tiu th cng nghip v hng trm c s u t nc ngoi, hu ht cc c s sn xut u cha c h thng x l kh thi hon chnh. Theo thng k cha y th lng kh nhim hng nm
thi ra 1017 tn bi, 30.580 tn SO2, 390 tn SO3, 1.948.500 tn CO2, 260 tn CO, 7.554 tn NO2, 137 tn hydrocarbon, 78 tn aldehyde. Ton b cc nh my thuc cng ti Thp min Nam hng nm thi vo mi trng t 2.840 tn n 4.600 tn bi v 994-1.420 tn CO. Ti thnh ph H Ch Minh vi s xe trn 900 ngn chic, hng thng tng 1200 chic cng hng triu lt xe t cc tnh v thnh ph tiu th khong 2-19 ngn tn xng, 190.000 tn CO, 12.000 ngn tn CO2, 13.200 tn hydrate carbon, 160 tn aldehyde. Tnh trn ton th gii, ring cc phng tin c gii thi ra 60% kh CO, 42% NO2 v 40% hydrate carbon, 13% mui v 3% SO2. Ring giao thng vn ti thi ra 18% tng lng CO 2 trn th gii tng ng 700 triu tn carbon mi nm (Cu ly tri t UNEP, 1992) Theo bo co ca vin Quan st Th gii, 1992, tc cht thi ca cc phng tin c gii M: CO l 60,3 triu tn, HC: 14,1 triu tn, SO2: 16,6 triu tn, NO2: 18,2 triu tn, SPM: 4,1 triu tn. Giao thng cng pht trin cng tng s nhim. Ngoi ra tu ha, tu thy chy bng than hay xng du u gy nhim mi trng tng t nh xe t. My bay cng l ngun gy nhim bi, hi c hi v ting n, c bit my bay siu m cn thi ra nitrooxide gy h hi tng ozone l tm l chn tia cc tm cho tri t. Ngun nhim khng kh t nng nghip to ra khong 15% tng s cc cht kh nhim gy bc x to nn hiu ng nh knh. Trong CO2 c to thnh do vic t rng lm nng ry v do ha hon. Trong thp k qua c 1.700.000km2/nm rng nhit i b tn ph m hu qu lng CO2 tng ln nhiu. Nhng cnh ng la min t t gii phng kh methan t cc qu trnh phn gii cht hu c. Ngun sn sinh cc cht ny ng k t cc trang tri chn nui v t cc ng rc khng x l ng k thut. Cc cht kh ny gy nhim mi trng lm tng hiu ng nh knh v ph hy tng ozone. Ngoi ra cc cuc chin tranh cng lm nhim mi trng kh rt cao. Lng bom n gy nhng tai ha v ngi v c th thng k, nhng nhng tai ha v mi trng sinh thi th khng th thng k v kh khc phc hu qu. Vit Nam trong thi k chin tranh chng M c hng triu tn bom, hng chc sn bay d chin vng rng ph i sinh cnh, min Nam 72 triu lt ha cht dit c, 44 triu lt cht c mu da cam ri xung tn ph rng v a vo mi trng cc cht c hi khc. Ngi dn vng v tuyn th 17 trong thi k M nh ph c lit, hng vn tn bom, n i bc ri vo qu hng h, thng xuyn b khi thuc sng quyn vi bi t m mch. Nhng ngi dn y vn kin
cng anh dng chng tr my bay, bom n ca quc M v h vn vui v ni a mt cch d dm: Qu ti cc nam ca min Bc, cc bc ca min Nam nn cc i l cc. Cuc chin tranh vng vnh 1991, hn 570 ging du b t chy, trung bnh mi ngy c khong 120 triu la M bin thnh khi en. Theo tnh ton trong thi gian xy ra cuc chin c khong 3 triu tn kh carbon, CO2, carbon hydroxide v cc ha cht c hi khc sn sinh ra t cc ging du b chy lm nhim nghim trng vng tri vng vnh v cc khu vc khc. Do du m chy khng ht nn trong khi du c nhiu cht c hi nh kh carbon, hydrocarbon, nh hng n sc khe con ngi. Khp mi ni th Kuwait u sc mi du chy, nhiu ngi b ho v nhc u. Trong nm 1991 Takzan xut hin nhiu trn ma acid u l ma acid qu mc bnh thng. Kh quyn b nhim khng nhng trc tip e da sc khe con ngi m cn nh hng rt ln n kh hu. Hin nay ton cu ang lo ngi v nhng mi e da ln lin quan n s nhim kh quyn l ma acid, hiu ng nh knh v thng tng ozone. * nhim mi trng t: nhim mi trng t l s xm nhp hoc a vo nhng cht l c kh nng tc hi n sinh vt v mi trng trong cc mi trng vi liu lng trn mc cho php . Cc ngun gy nhim mi trng t - T t nhin: Cc loi nhim do s c ca mi trng nhng vng khc nhau. Chng hn, ni la l hin tng s c gy nh hng n mi trng t, nt t cng nh hng n mi trng t, s tun hon nc vi mt tc qu ln t ngt cng l mt hin tng gy st l. - T con ngi: y l ngyn nhn ln nht + T hot ng cng nghip: thi vo mi trng khng kh mt lng bi trong c cha nhng dng nhim nh c gc acid (Cl-, SO4--) khi kt hp vi nc to thnh m my acid ri xung lm nhim mi trng t, pH ca t thay i. Bi thi c nhng kim loi nng vo khng kh nh Pb, Zn, As, ... nh trng lc ca chng ri xung t lm nhim mi trng t. Ngoi ra hot ng cng nghip cn thi ra mi trng t ngun nc bn ri thm thu, khuch tn vo mi trng t gy nhim. + Hot ng nng nghip: gy nhim theo nhng hng khc nhau nh: s dng phn bn ha hc, ngun thuc bo v thc vt.
+ Sinh hot: ch yu l rc thi, bnh qun 1 ngi/1ngy thi 4 kg rc thi. Cc rc thi ny thay i theo thi gian v mc sng ca tng vng, c bit khi cng nghip ha-hin i ha pht trin th ngun rc thi cng phong ph v kh phn hy. * nhim mi trng nc: L s a cc cht l vo trong ngun nc do cc hot ng ca t nhin, con ngi qu tiu chun cho php gy nh hng n i sng, sc khe ca con ngi. Cc ngun gy nhim mi trng nc: Ngun gc gy nhim c th l t nhin hoc nhn to. S nhim c ngun gc t nhin c th do ma, tuyt tan, bo t, ni la,..N rt nghim trng nhng xy ra khng thng xuyn v khng phi l nguyn nhn chnh gy suy thoi cht lng nc ton cu. S nhim nhn to ch yu l do nc thi sinh hot, nc thi cng nghip, giao thng vn ti, phn bn v cc ha cht bo v thc vt. Da vo ngun gc pht sinh m ngi ta phn ra thnh cc loi nc thi: nc thi sinh hot, nc thi cng nghip, nc ma chy trn, nc sng, h, bin b nhim do cc yu t t nhin. c tnh cc loi nc thi: - Nc thi sinh hot: Nc thi sinh hot l nc thi ra t cc h gia nh, bnh vin, khch sn, trng hc, c quan,...Loi nc thi ny cha hm lng cc cht hu c khng bn vng kh cao, d b phn hy sinh hc. Cc cht ny khc nhau, ty thuc vo iu kin sng khc nhau. M l 380 - 500 lt/ngi/ngy m. - Nc thi cng nghip: S tng nhanh v cng nghip lm tng nhu cu v nc v lng nc thi t cng tng ln. c bit i vi cc ngnh ch bin thc phm, giy, ha cht, du m, than v luyn kim. Nm ngnh ny tiu th ngt 90% tng lng nc trong sn xut. Chng hn phi s dng 15 lt nc sn xut 1 lt bia, 200 m 3 nc sn xut 1 lt du lc, 300 m3 nc sn xut 1 tn giy. - Nc chy trn mt t: Do nc ma hoc do thot nc t ng rung cng l ngun gy nhim nc sng, h, bin. Nc ra tri ng rung c th cun theo cc cht rn, ha cht bo v thc vt, phn bn. Nc ra tri qua khu dn c, ng ph, c s sn xut cng nghip c th lm nhim ngun nc do cc cht rn, du m, ha cht, vi sinh vt,... - Nc sng, h b nhim do cc yu t t nhin:
17 1 CNSH vi vic bo v mi trng Trng Vn Lung
Nc sng, h b nhim mn vng ven bin c th chuyn nc mn vo cc vng su trong ni a. Nc sng, knh rch b nhim phn c th chuyn acid, st, nhm,... n cc vng khc v gy suy gim cht lng nc ca vng b tc ng. TI LIU THAM KHO 1. Hong Hu, 1996. X l nc thi. Nxb Xy dng. H Ni. 2. Trn Vn Nhn, Ng Th Nga, 1999. Gio trnh cng ngh x l nc thi. Nxb Khoa hc K thut. H Ni. 3. Trn Cm Vn, Bch Phng Lan, 1995. Cng ngh vi sinh v bo v mi trung. Nxb Khoa hc & K thut. Trung tm Giao lu quc t v Vn ha Gio dc v Khoa hc (CCES), H Ni.
CNSH vi vic bo v mi trng
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Chng IX
Vai tr ca cng ngh sinh hc trong vic x l mi trng

1. Nhng nghin cu trong vic chng nhim mi trng
Ngy nay, x l cht thi ngi ta phi hp cc bin php c hc, vt l (lc, nghin, sa lng...), bin php ha hc (phn hy, dit sinh vt hi...) kt hp vi bin php sinh hc thnh qui trnh x l hin i v mang tnh trit nht. Ty theo loi cht thi khc nhau m cc cng on, trnh t x l c khc nhau loi tr cc cht, thnh phn khng mong mun. Trong qui trnh ny, cng on x l sinh hc li dng c tnh sinh vt bin i thnh phn cht thi. C th tm lc qui trnh nh sau: - S dng cc vi sinh vt phn gii ym kh phn gii cc cht thi c ngun gc hu c. - S dng cc vi sinh vt, enzyme phn gii hiu kh cc cht thi, bin cht thi thnh cht c th s dng c. - S dng vi sinh vt hp thu cc cht kh tan, bin chng thnh cc sn phm khc, bin i tnh cht ca cht thi. - Dng thc vt thy sinh hoc trng hoa mu di dng thy canh ht cc cht bn ha tan trong mi trng nh NH 4+, NO3-, PO43dng lm thc n ng ha trong c th thc vt. Lc ny mi trng mi sch thc s.
2. Ch bin v s dng cht thi trong mi trng

Mt s qui trnh x l cht thi Qui trnh x l cht thi c in T xa con ngi bit x l cht thi bng cc bin php ht sc n gin, th s nh t, chn lp cc cht thi rn, vt rc thi trc khi cho thi ra sng, ao h. c bit i vi cht thi t chn nui (phn) ngi ta vi rm r v dng lm phn bn cho cy trng... Do mt dn c cha cao, thm vo l nn sn xut th s nn cht thi khng nhiu, hn na ch yu l cc dng d phn gii nn cht thi c mt thong, thi gian phn gii, chuyn ha vo chu trnh vt cht. Qu trnh x l kh n gin, ch yu da vo cc sinh vt c sn trong t nhin, vn rc thi cha mang tnh x hi.
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Qui trnh x l cht thi hin i Cng vi s pht trin ca nn vn minh v tc pht trin ca loi ngi, mi trng ngy cng b nhim nghim trng. Nh vy, nhim mi trng ang l vn quan tm ca ton nhn loi. Con ngi phi gii quyt vn nhim mi trng vi cc qui trnh, cng on phc tp, c s tham gia ca nhiu lnh vc khoa hc: vt l, ha l, sinh hc... Trong nhng gii php khoa hc th cng ngh sinh hc (CNSH) ng gp vai tr ch o, quyt nh nht. CNSH ngy nay tr thnh cng c c lc con ngi bo v c mi trng sng khc ca hnh tinh. CNSH c p dng cho nhiu lnh vc khc nhau bo v mi trng. Dng cy c nh m bo v, ci to t t trng, i trc l t ai d b thoi ha do xi mn, ra tri. chng s thoi ha ca t y th bin php hng u l ph xanh cc vng t . Tuy nhin t y ngho cht dinh dng v vng t ny thng rt chua. Do bin php u tin l phi tm ra loi cy thch hp vi vng t. Theo nhiu nghin cu th cc cy h u thch hp vi vng t ny, chng c kh nng ci to t rt tt. Chng b sung m cho t ci to khu h vi sinh vt quanh r gp phn thc y qu trnh to mn trong t, gi nc hn ch xi mn. *Phng php trng cy b u: - Chn ging: do iu kin khc nghit ca vng t trng, i trc, mun cho cy pht trin tt trc ht cn chn ging thch hp. + Chn lc ging no pht trin tt trn t trng, i trc trong s tp on cc ging cy b u. + Lai to to ra cc ging cy u thch hp vi vng t . + Hoc dng cng ngh gene to ra ging cy chu hn, cn t cht dinh dng, chu chua... thch ng vi iu kin khc nghit vng i trc m vn cho nng sut tt. - Nhn ging: dng phng php nui cy m t bo nhn ging cy c chn, p ng nhu cu trng nhng din tch ln. - Trng cy b u ln vng i trc. Ty theo iu kin t ai, kh hu v nhu cu ca tng ging cy m c bin php k thut hp l. * Nhn ging vi khun Rhizobium Song song vi vic chn ging cy b u, nhn ging v trng cy cn tin hnh vic chn lc, nhn ging vi khun Rhizobium thch hp vi cy b u em trng ci to t.
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Dng vi khun Rhizobium sn xut phn vi sinh (nitragin) bn cho cy. C th dng nitragin tm ht hay bn vo t trc khi trng cy b u. To kh nng c nh m: to nn cc dng cy c kh nng c nh m t cung cp m. Dng cng ngh gene chuyn gene sn xut enzyme nitrogenase (gi l gene nif) t vi khun c nh N2 sang cho E.coli to cho E.coli c kh nng c nh N2, ri s dng E.coli ny cho cy trng nh l ngun cung cp N2 cho cy Dng phn sinh hc chng nhim Phn ha hc c vai tr ln trong vic nng cao nng sut cy trng. Tuy nhin nu lm dng phn ha hc qu th s dn n nhng tc hi nghim trng, nht l gy nhim mi trng. trnh nhim mi trng bin php c hiu qu nht l s dng phn bn sinh hc thay th cho phn ha hc. Bi v phn sinh hc c vai tr ln trong vic nng cao nng sut cy trng nhng khng gy nhim mi trng, gp phn xy dng mt nn nng nghip sinh thi. Phn vi sinh l dng c nhiu u im. CNSH to ra nhiu dng phn vi sinh c gi tr s dng cao, vi hiu qu khc nhau ph hp vi cc loi cy trng khc nhau, cc loi t trng khc nhau. Ph bin nht hin nay l phn sinh hc nitragin v Azotobacter, ngoi ra cn c phn ln sinh hc. Dng phng php sinh hc bo v thc vt Thuc tr su ha hc gy nhiu tc hi cho mi trng nn vic s dng bin php sinh hc bo v thc vt c u im rt ln trong bo v mi trng, bo v tnh a dng sinh hc, bo m s cn bng sinh thi. Nhng bin php sinh hc bo v thc vt gm: - Dng vi sinh vt dit cn trng gy bnh: C 3 nhm c kh nng gy bnh cho cn trng l vi khun, vi nm v virus. Trong nhm vi khun ngi ta thng dng Baccilus thuringiensis to ch phm tr su sinh hc. - Dng vi sinh vt dit vi sinh vt gy bnh: Chng vi sinh vt thng c s dng to ra thuc tr su sinh hc l x khun phn lp t t. Sau khi chn c chng thch hp tin hnh nui cy nhn ging trong ni ln men. Dch nui cy chnh l ch phm thuc tr su sinh hc th. T ch phm c th to ra thuc tr su dng nc hoc dng bt s dng. - Dng cn trng, ng vt khc dit cn trng gy bnh.
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Nguyn tc chung ca cc bin php trn l s dng tnh i khng sinh hc gia cc nhm sinh vt vi nhau tiu dit cc sinh vt gy bnh cho cy. Khi s dng thuc tr su, khng ch cn trng gy hi b tiu dit m cc loi i khng vi chng cng b tiu dit theo lm cho yu t t iu chnh ca h sinh thi t nhin b mt i, h sinh thi tip tc mt cn bng ngy cng nghim trng thm. Cc phng php sinh hc bo v thc vt l nhm vo vic phc hi nhng loi i khng vi cn trng gy hi tiu dit cc loi cn trng gy hi m khng lm nhim mi trng. Cng ngh x l rc thi Rc l mt nguyn nhn nghim trng gy nhim mi trng nh mi trng nc, t, khng kh. Rc thi trong mi trng hin nay ch yu l rc thi sinh hot v rc thi cng nghip. Thnh phn rc rt a dng. Rc sinh hot ch yu l giy loi, nha, tng hp thy tinh cao su, ph thi t thc n... Thnh phn ha hc phc tp gm cht v c v cht hu c... kh phn hy cn rc trong cng nghip gm nhiu loi ph thi nh kim loi, thy tinh, nha, giy... c thnh phn l ha khc nhau nn phng php x l chng cng rt phc tp. Vic x l rc thi m khng lm nh hng n mi sinh khc nh kh, nc... l mt vn ln. Trc y, phn ln rc ca cc thnh ph, khu vc dn c... c tp trung li ri chn hoc t i. C 2 phng php ny u lm cho nhim li mi trng, l phng php x l chm li t hiu qu trong thi i hin nay do s pht trin ca khoa hc hin i. S pht trin ca khoa hc cng ngh gp phn rt ln vo cng cuc bo v mi trng xanh sch p nh cng ngh ha cht... nhng nhng cng ngh va x l nhng li va lm nhim ngha l cng vic x l mi trng bng phng php l ha cha em li kt qu ti u. Ni nh vy l ta thy c ng dng v vai tr rt ln ca cng ngh sinh hc trong x l rc thi. X l mi trng ni chung (rc thi ni ring) bng phng php sinh hc em li hiu qu cao nht, va khng lm nhim mi trng tr li, va c hiu qu kinh t cao. C nhiu phng php x l ph thi khc nhau ang c s dng... Bn cht ca cc phng php th hin trong 4 ni dung ch yu sau: - Phn hy s cp lm thay i bn cht ha hc ca cc cht x l. - Phn hy sinh hc s cp lm lm thay i tnh cht ca cht x l. - Phn hy sinh hc ring bit bin ph thi hu c thnh sn phm v c.
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- Ht cc cht v c cho mi trng tr nn sch hn nh thc vt trong qu trnh ng ha ca mnh to nn cc sn phm hu ch. Cc qu trnh trn thc hin c nh c s tham gia tch cc ca cc nhm vi sinh vt v thc vt thch hp. Do thnh phn ch yu ca ph thi l cellulose cho nn cc vi sinh vt c kh nng phn gii cellulose c s dng vo vic x l ph thi rt hiu qu... C nh vy, ty thuc vo thnh phn ca tng loi rc m ta chn v s dng vi sinh vt cho ph hp. * Vic s dng vi sinh vt phn hy rc c tin hnh qua cc bc sau: Phn lp vi sinh vt phn hy rc. Chn chng thch hp, nng cao hot tnh phn hy rc. Th nghim kh nng phn hy rc. Ch to ch phm. Cc ch phm khi em s dng phi ty thuc vo loi rc, ty chng vi sinh vt m c th thc hin cc phng php phn hy khc nhau: phn hy k kh, phn hy hiu kh hay phn hy hiu kh-k kh phi hp. Rc sau khi phn hy c dng lm phn bn. cc nc tin tin ngi ta tin hnh x l rc thi theo cc phng php hin i bng cc thit b hin i. Cc phng php cng da vo kh nng phn hy rc ca cc nhm vi sinh vt c tin hnh trong cc thit b khp kn nn khng nh hng n mi trng. C 2 phng php x l rc ch yu theo cc cng ngh tin tin ny: - X l ph thi bng con ng hiu kh, con ng ny c 3 cng ngh ang c s dng: * X l bng mng lc thm thu. * X l bng bung hot tnh. * X l bng lp men gi. y l mt cng ngh s dng rng ri vo khong 2 thp k gn y, c bit cc nc ang pht trin nh Trung Quc,Vit Nam. + X l ph thi bng con ng ym kh l qu trnh x l ph thi s dng cc vi sinh vt ym kh phn hy cht hu c trong ph thi thnh kh methan. y l hng va x l ph thi va to ra biogas s dng trong sinh hot. Phng php ny c s dng rng ri n , chi ph t v tit kim v kinh t. + Cc ph thi cng nghip c chia thnh 2 nhm: - Cc ph thi c lin quan n sinh hc nh cng nghip sa ng, thc phm...
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- Cc ph thi khng lin quan n sinh hc nh cng nghip nng, c kh... i vi loi ny thng x l bng phng php l ha. i vi ph thi c lin quan n sinh hc th ty theo loi ph thi m c phng php x l thch hp: + X l ph thi ngnh cng nghip sa Huyt thanh sa l ph thi ch yu ca ngnh cng nghip sa. Vi loi ph thi ny ngi ta thng c c lm thc n gia sc hoc cho ln men Lactobacillus bulgaricus thu sinh khi sinh vt giu protein lm thc n cho gia sc. + X l ph thi ngnh cng nghip giy Phn ph thi nh my ph thuc phng php thy phn nguyn liu lm giy. Nu x l bng kim th ph thi cha nhiu ligin kh phn gii v cc acid hu c. Ph thi ny kh x l bng phng php sinh hc nn thng t tn dng nng lng. Nu x l nguyn liu bng acid th ph thi cha 60% lignin, cc loi ng chim 30% v cc cht khc nh acetic, methan... cc loi ph thi ny dng phng php ln men to nn sinh khi giu protein. + X l ph thi ngnh cng nghip nhum Ph thi cha nhiu mu v sc t khc nhau. Ngoi ra cn cha nhiu cht c nn ch yu dng phng php ha hc. + X l nhim du m Du m l nguyn nhn gy nhim mi trng nghim trng trong khai thc, vn chuyn v s dng. x l nhim do du m ngi ta thng s dng cc vi sinh vt s dng du lm thc n x l. X l nc thi bng CNSH C nhiu phng php x l nc thi, mi phng php u c u, nhc im ca n. - Phng php ha hc: dng cht kt ta, lc kt hp vi s phn hy ha hc. - Phng php vt l: dng nhit dit trng v phn hy. - Phng php ha l: trao i ion, hp th. - Phng php sinh hc: s dng vi sinh vt v thc vt. Trong cc phng php trn phng php sinh hc ngy cng c s dng rng ri v phng php ny c nhiu u im hn cc phng php khc: - Phn hy cc hp cht hu c trong nc thi nhanh, trit m khng gy nhim mi trng. - To ra c mt s sn phm c ch s dng trong cng nghip v sinh hot (nh biogas, ethanol...), trong nng nghip (phn bn).
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- Thit b n gin, phng php d lm, chi ph tn km t hn cc phng php khc. Nguyn tc c bn ca phng php sinh hc x l nc thi l dng h vi sinh vt phn hy cc cht c trong nc thi to nn cc sn phm khng c hi cho mi trng. Cc sn phm ca qu trnh phn gii nc thi do vi sinh vt c th c s dng trong nhiu lnh vc ca sn xut i sng nh nh to ra biogas, to protein trong sinh khi vi sinh vt lm thc n gia sc... H vi sinh vt tham gia trong x l nc thi c nhiu loi khc nhau nh nm men, nm mc, x khun. Ty theo h vi sinh vt s dng m c phng php x l thch hp theo hai hng: x l ym kh v x l hiu kh hay x l t nhin. *Phng php hiu kh x l nc thi y l phng php x l nc thi trong iu kin c O 2 tham gia. - S dng bn hot ng x l. Bn hot ng l mt hn hp dng lng st nh bn trong cha nhiu vi sinh vt hiu kh (vi sinh vt chim 70% khi lng ca bn). Bn hot ng c kh nng v c ha mnh cc hp cht hu c c trong nc thi. H sinh vt dng trong bn hot ng gm nhiu nhm khc nhau, thng gp l Actinomyces, Bacillus, Bacterium, Pseudomonas, Micrococus... Cho bn hot ng chy vo b x l nc thi. Ti b x l cc vi sinh vt s tin hnh phn hy cc cht c trong nc thi trong iu kin khng kh lun lun c sc vo b. Sau khi qua b x l, tin hnh qu trnh lng hn hp lc phn nc sch v tch phn cn b. Phn nc sch s c a ra s dng li theo cc mc ch ring. *Dng b x l hiu kh: bn hot ng cha sn trong b, cho nc thi chy vo b, khuy sc kh. Nc sau khi c x l ly ra bng van v em s dng li, ngoi ra theo phng php ny cn thu c biogas. *Dng b lc sinh hc: ngi ta cho vo b vt liu xp, mang vi sinh vt. Vt liu xp c th l mnh gm, s, dm, si, polymer... vi xp cao. Nc thi phun t trn xung, khng kh thi t di ln. Nc x l s b a sang b lng tch. *Phng php x l ym kh nc thi Nguyn tc ca phng php ny l s dng cc vi sinh vt ym kh x l nc thi trong iu kin khng c O2.
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Mt trong nhng bin php ym kh x l nc thi l dng b methan x l. Phng php ym kh ch yu dng cho loi nc thi c nng cht hu c cao, c nhiu cn, cht x thng dng c qunh nh bn. Qu trnh lm sch nc thi tin hnh trong b kn m bo iu kin ym kh. Cho nc thi vo b vi sinh vt phn gii ym kh s tin hnh qu trnh phn gii cht hu c c trong nc thi. Sn phm ca qu trnh phn gii ny l biogas chim 50-80%. Ngoi ra cn c mt s cht khc nh benzol, toluel... H vi sinh vt ln men ym kh thng c sn trong nc thi. Tuy nhin tng nhanh tc phn gii, nng cao nng sut hot ng ca cc b methan, c th phn lp, nui cy cc vi sinh vt thch hp cung cp thm cho b. Cc nhm vi sinh vt thng gp trong qu trnh phn hy ny l Methanococcus, Methanobacterium, Methanosarcina... Do ngha ca b methan m hin nay phng php ny ang c ph bin rng ri t qui m nh h gia nh cho n qui m cng nghip va x l nc thi gii quyt vn nhim mi trng va to ra kh biogas cung cp cho sinh hot. c bit qui m h gia nh th phng php ny rt tin li v c hiu qu cao nn c ng dng kh rng ri trong nhn dn. X l bng thc vt thy sinh Ngoi vic x l nc thi bng cc loi vi khun hiu kh, ym kh nhm loi bt cc lng cht hu c trong nc thi, ngi ta cn s dng mt s loi to, bo v vi sinh vt khc. C s khoa hc ca gii php ny l mt loi to v vi sinh vt c kh nng tch ly cc kim loi c nng cao nh: Fe, Cu, Zn, Mo,.. v cng c th tch ly nhng cht khc. X l nc thi bng to nh: Theo Lalierte v cng s (1994), vic dng to nh x l nc thi c nhng u th ring bit. Nc thi t h thng x l th cp, tuy c hm lng BOD v COD thp, nhng hm lng cc cht v c (NH 4+, PO43-, NO3-,...) cn rt cao. To nh s hp thu cht ny, ng thi to ra lng oxygen cn thit h tr c lc cho qu trnh oxyha sinh hc ca vi khun. Nui trng to nh trong mi trng nc thi chnh l tn dng u im ca chng v kh nng bin i c hiu qu cc cht dinh dng trong nc thi thnh ngun protein cao cp. ny c thc nghim chng minh nhiu ni trn th gii. Cc nh khoa hc Canada v Anh nui to Spirulina maxima trong nc thi ca x nghip x l nc thi sinh hot. Nc phn gia sc c cc nh khoa hc Israel v Bungarie
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dng nui to nh kh tt. Vit Nam, vic th nghim nui trng Spirulina platensis bng nc thi hm biogas khng ch l bin php m rng sn xut v h gi thnh sn phm m cn gii quyt mt khu quan trng trong x l nc thi lm lnh mnh mi trng sinh thi nng thn. X l nc thi bng thc vt thy sinh: C s khoa hc ca vic dng thc vt thy sinh trong x l nc thi l kh nng hp th cc cht dinh dng c trong nc thi ca chnh cc loi thc vt ny. Mt khc, quan trng hn l kh nng phn gii v ng ha cc cht bn trong nc thi ca vi sinh vt sng bm trn thc vt thy sinh. Thc vt thy sinh nh l mt gi th thch hp cho vi sinh vt chng tin hnh phn gii cc hp cht hu c trong nc. Thn v r ca thc vt thy sinh l gi th cho nhiu loi vi sinh vt pht trin. Hn na, s c mt ca thy sinh vt trong cc h n nh sinh hc hn ch v kim sot s pht trin ca cc loi to do lm gim kh nng xuyn thu ca nh sng v cnh tranh v dinh dng. V d: cng trnh th nghim s dng bo sen (Eichhornia crassipes) lm sach mi trng nc thi sinh hot ti hi Ph An v Ph Thng, huyn Ph Vang, tnh Tha Thin - Hu (2002).
Hiu qu x l cc cht nhim ca bo sen ti cc thi im nui
Cc thng s Hiu qu loi b cht bn sau 10 ngy (%) Ph An Ph Thng Hiu qu loi b cht bn sau 15 ngy (%) Ph An Ph Thng
NO3PO43NH4+ COD BOD5 Coliform
64.16 79.49 56.14 55.74 55 80.83
64.08 78.38 66.94 56.67 55.6 86.36
70.63 84.95 64.39 71.52 65 90
72.02 82.13 73.24 63.78 63.9 87.27
i vi th nghim ny, cc tc gi Trng Vn Lung, Hong Th Tuyt Ngc tin hnh th nghim vi nhiu khay mu c cc su khc nhau: 25cm, 35cm, 45cm. bng trn, chng ti ch trch s liu in hnh i vi khay mu su 35cm. *Phng php t nhin x l nc thi Vi nhng loi nc thi c nng cht nhim bn khng cao, khng cha nhiu cht c hi cho con ngi th thng dng phng php x l t nhin.
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Nc t nh my, khu dn c... cho chy thm qua lp t dy khong 2 m, hay chy qua lp t xp, ct... Nc thi i qua cc lp nh h vi sinh vt c sn y, cc cht bn s c phn hy lm cho nc tr nn sch hn. y l qu trnh t lm sch nn d lm, t tn km, c th x l nc thi trn qui m ln. Nc sau khi x l c th dng trong nng nghip, dn ra ao, h, sng... ha vo ngun nc chung ca t nhin. Mt s cng trnh nghin cu, ng dng CNSH 1. Chng trnh biogas ca Trung Quc ci tin vic sn xut v tiu dng nng lng cho gia nh m cn ngn cn vic gy nhim. Cht thi ca qu trnh lm biogas c dng lm phn bn, trng nm, lm thc n cho c, gp phn lm sch sn phm ph v loi b cc ph phm. Protein n bo (SPC - single cell protein), dng c cht l hydrate carbon, r ng, b thi ca nh my bt giy nui cy sng cc nm men Sacchromyces, Candida, Mycoprotein, cc loi to n bo Chlorella, Scenedesmus, Spirulina... Cc nc tin hnh l Thy S (Cng ti Envirocom Lid, Vancower), Phn Lan (G. A. Serlacchus), Cuba. 2. K thut nui to trn nc thi pht trin r v c nhng ci tin quan trng, lm cho nc trong v sn sinh mt lng sinh khi giu protein v nguyn t vi lng. Mt vi loi to thuc chi Pseudomonas c nhng enzyme oxyhakh hay hydroxyl ha phn hy c mt s ln phn t hydrocarbon hoc cc hp cht thm rt c (benzyl, toluen, xylen...). Nm 1979, Chakrabarky i hc Illinois chn c mt chng vi khun c kh nng pht trin mau chng trn du th. Sau mi thng nui cy, t l tng trng ca chng tng ln ng k v 2,4,5T (2,4,5 acid trichlorophenolacetic) c tiu i trong vng vi ngy thm ch c nng rt cao. K thut ti t hp DNA thng cung cp kh nng to ra nhng chng vi khun c th ph hy v hp thu mt s ln cc cht do cng nghip ha cht sn xut ra thng thng khng phn gii c bng con ng sinh hc, v d Nht Bn, Furukuwa v cng s tch c mt chng x khun Nocardia c th phn hy cao su ch trong mt tun. Cht thi rn X l rc thi bng giun Giun n l v thi ra cc cht hu c lm t ti xp. Cc nh khoa hc Vit Nam th nghim thnh cng phng php nui giun bng rc thi, nhm gii quyt nn nhim mi trng do rc gy ra, ng thi
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cung cp thc n cho gia sc. Loi giun ny c nhp t Philippines, c u im l d nui, sinh sn nhanh, thch nghi tt vi kh hu nc ta. Hunh Th Kim Hi, thuc vin Sinh thi v Ti nguyn sinh vt, nghin cu kinh nghim dn gian, kt hp vi cc kin thc khoa hc hin i cho ra i mt qui trnh x l rc thi nh giun t Phillipines. Loi giun ny c tn khoa hc l Perionyx excavalus, c th tiu ha cht thi rt tt. Theo tnh ton, phn hy 1 tn rc hu c trong mt nm, ngui ta cn khong 1.000 con giun ging v cc th h con chu ca chng. Hin ti, ti nghin cu c ng dng cho vic x l rc thi cc thnh ph ln. Trn thc t, vic nui giun t x l nhim mi trng c nhn dn ta p dng t lu. Kinh nghim ny c ph bin rng ri nht H ng. Nhn dn y thng lm chung g pha trn v nui giun t pha di, v phn do g thi ra l ngun thc n tt cho giun t. Mt khc nh giun n t, tiu ha v thi ra cht hu c, m sau mt thi gian, t pha di chung g s ti xp, rt tt cho cy trng. Khi , ngi ta li chuyn chung g ra ch khc, c nh vy... Chu trnh khp kn ny khin cho vic nui gia cm khng gy nhim mi trng. Kh mi rc bng ch phm sinh hc Mt lt ch phm sinh hc Odor removal (Uyama enzyme) ca Cng ti TNHH Thi Dng (Long An) c th x l 20 tn rc. Loi ch phm c kh nng kh mi hi ca rc hn 60% trong ngy u tin v gim dn n hn 80% trong cc ngy tip theo. Mi hi thi trong nc r rc cng gim hn 80% v khng ti nhim, mt rui gim hn 90%. y l bn nh gi kt qu kho nghim ca hai trng i hc Khoa hc T nhin v Bch khoa thnh ph H Ch Minh v ng dng ch phm ny ti cc trm trung chuyn rc, bi rc v nc r rc. Bao gi t phn hy sinh hc (kt hp phng php ha hc v sinh hc) L Trung Sn v Trnh L Hng, ging vin khoa Ha, trng i hc Khoa hc T nhin H Ni, ch to thnh cng bao gi t phn hy sinh hc. Bao gi c lm t hn hp tinh bt (bt ao v bt sn) trn vi cc dung dch protein v cc cht do ha, cht ph gia to thnh hn hp blend. Tri hn hp ny ln b mt ri gia nhit, vt liu c to ra sau khi sy l mt mng mng c dy t 0,15 - 0,35 mm.
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Mng vt liu ny c dng ch to cc chu, ti trng cy t phn hy, cc ung nc, giy gi ko, a thc n dng mt ln....Vt liu ny c gi t 1.000 - 1.200 ng/m2. Cc kt qu th nghim cho thy vt liu phn r trong nc sau vi ngy, trong t sau vi tun. Nhng sn phm lm t vt liu ny s hn ch nhim mi trng. Cng ngh vi sinh trong sn xut ch phm sinh hc BZT Gii thch v vi khun: Qu trnh n nh vi khun lm vic nh th no? Vi khun sinh sn bi mt qu trnh gi l nhn i t bo mi sinh ra bng cch chia i t t bo m. Mt s c th sinh sn rt nhanh nhng iu kin thch hp. Nu c thc phm, m thch hp ng nhit vi sinh c th bt u sinh sn trong vng cha n hai mi pht, ch trong tm gi mt t bo ban u c th nhn ln thnh 17 triu vi khun mi. Vi khun chn lc, c nui cy trong mt mi cht. Di s gim st, vi khun sau khi thanh lc c chuyn qua mt bn ln men sinh hc 250 lt sinh trng trong vng 20 gi. Sau khi kim tra cht lng v thun khit, vi khun c a sang bn ln men kn v trng c dung tch 5.000 lt bt u giai on sn xut. Di iu kin pH thch hp, dung dch ng v trng v oxygen c a vo nui vi khun. Trong sut qu trnh ny, sn phm c ly mu theo di s v trng v cc thng s tng trng. Trong vng 24 gi, s thu hoch vi khun v c c bng mt my li tm cc nh. Vi khun c c c bc li bng cht keo -glucan bi mt qui trnh c cp bng sng ch. Qui trnh ny gip vi khun chng li m duy tr s sng trong thi gian bo qun hoc trn vi cht mang. Sau sn phm c lm lnh nhanh trong h thng lnh lng trc khi a vo sy nhit -400C trong h thng phng lnh ln. Qui trnh sy lnh hai bc ny, trong iu kin m di 5%, bo m t l sng st ca vi khun cao v sn sng cho giai on cu l kim tra m bo cc dng vi khun khng nhim khun Salmonella. Chuyn i BOD v oxygen BOD (nhu cu oxygen sinh hc s oxyha sinh hc) l khi lng oxygen ha tan c tiu th bi cc vi khun trong qu trnh oxyha cc cht hu c v v c. Th nghim BOD c xy dng nhm ngn nga hu qu ca nc thi ti ngun nc v xc nh kh nng tiu ha cc cht hu c ca chng.
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S chuyn i oxygen ln trong nc l mt vn phc tp v c nhiu nh khoa hc b ra c cuc i nghin cu. Vic cho rng oxygen chuyn i bng nhng bt kh ln ch ng trong mt vi trng hp ch yu trong vic ln men khi m protein bao ph b mt bt kh. chnh l nguyn nhn ti sao phi s dng my p kh v bm kh trong qu trnh ln men. ng dng cng ngh vi sinh vt trong sn xut phn bn hu c sinh hc S dng nhng chng vi sinh vt c tuyn chn nh hng sn xut phn hu c vi sinh va lm rt ngn thi gian chuyn ha cht hu c sang dng mn li va nng cao cht lng v hiu qu phn bn. rc vi ch phm Micromix 3 rt ngn 14 ngy so vi rc thng thng, cht lng mn trong sn phm tng. Tip theo b sung cc ch phm ging vi sinh vt hu ch cho cy trng to nn ch phm phn hu c- vi sinh cha 106 CFU/g tng loi vi sinh vt hu ch cho cy trng (c nh N2, phn gii phosphate, sinh cht kch thch sinh trng). Loi phn bn ny c tc dng tng nng sut cy trng, gim lng phn bn ha hc, tit kim u t sn xut v gim nhim mi trng t, nc v thc phm Seraphin - Cng ngh x l rc thi sinh hot ti Vit Nam Rc thi sinh hot ti Vit Nam, nht l ti cc thnh ph ln ch yu c x l th s bng cch vi ti cc bi chn lp, c nguy c gy nhim mi trng v ngun nc ngm. Cn c vo thc t , tp th cc nh khoa hc thuc cng ti c phn Cng ngh Mi trng xanh t nghin cu v pht trin thnh cng cng ngh x l rc mang tn Seraphin, ph hp vi c im rc thi Vit Nam l khng c phn loi t u ngun. Cng ngh Seraphin c nghin cu trong 5 nm v c ng dng cch y 18 thng di dng nh my x l rc th im Ninh Thun vi cng sut 150 tn/ngy. Chi ph xy dng l 20 t ng. C th tm tt qu trnh x l rc thi nh sau: Ban u rc t khu dn c c a ti nh my v xung nh tp kt ni c h thng phun vi sinh kh mi cng nh ozone dit vi sinh vt c hi. Tip n, bng ti s chuyn rc ti my x bng ph v mi loi bao gi. Rc tip tc i qua h thng tuyn t (ht st thp v cc kim loi khc) ri lt xung sng lng. Sng lng c nhim v tch cht thi mm, d phn hy, chuyn rc v c (k c bao nha) ti my v v rc hu c ti my ct. Trong qu trnh vn chuyn ny, mt chng vi sinh ASC c bit, c phun vo rc hu c nhm kh mi hi, lm chng phn hy nhanh v dit mt s
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tc nhn c hi. Sau , rc hu c c a vo bung trong thi gian 7-10 ngy. Bung c cha mt chng vi sinh khc lm rc phn hy nhanh cng nh tip tc kh vi khun. Rc bin thnh phn khi c a ra khi nh , ti h thng nghin v sng. Phn trn sng c b sung mt chng vi sinh c bit nhm ci to t v bn cho nhiu loi cy trng, thay th trn 50% phn ha hc. Phn di sng tip tc c a vo nh trong thi gian 7-10 ngy. Do lng rc v c kh ln nn cc nh khoa hc ti cng ti tip tc pht trin h thng x l ph thi tr v do, to ra mt dy chuyn x l rc khp kn. Ph thi tr v do i qua h thng sy kh v tch lc bi tro gch. Sn phm thu c giai on ny l ph thi do sch. Chng tip tc i qua t hp bm ct, phi trn, s ch, gia nhit bo tn ri qua h thng thit b nh hnh p lc cao. Thnh phm cui cng l ng cng panel, cc gia c nn mng, vn sn, cp pha, gch bloc... C 1 tn rc a vo nh my, thnh phm s l 300-350 kg seraphin (cht thi v c khng hy c) v 250 - 300kg phn vi sinh. Loi phn ny hin c bn trn th trng vi gi 500 ng/kg. Hin cng ti ang lm ch u t xy dng nh my x l rc Thy Phng ti thnh ph Hu vi cng xut 150 tn/ngy, chi ph xy dng 30 t ng. Theo d kin, nh my s i vo vn hnh trong thng 11/2005. Mt nh my khc mang tn ng Vinh ti thnh ph Vinh vi cng sut x l 200 tn/ngy cng s c hon tt vo thng 12/2005 vi chi ph xy dng khong 45 t. Chi ph xy dng mt nh my x l rc sinh hot s dng cng ngh seraphin r hn nhiu so vi cc gii php x l rc nhp ngoi. Nh vy, qua cc cng on tch lc - ti ch, cng ngh seraphin lm cho rc thi sinh hot c ch bin gn 100% tr thnh phn bn hu c vi sinh, vt liu xy dng, vt liu sn xut dn dng, vt liu cho cng nghip. Cc sn phm ny c c quan chc nng, trong c Tng cc Tiu chun o lng cht lng kim nh v nh gi l hon ton m bo v mt v sinh v thn thin mi trng. Vi cng ngh seraphin, Vit Nam c th xa b khong 52 bi rc ln, thu hi t bi rc s dng cho cc mc ch x hi tt p hn. Tuy nhin, to iu kin d dng hn trong khu x l rc thi sinh hot, cng ti V sinh Mi trng th ti cc tnh, thnh ph cn vn ng, hng dn ngi dn phn loi rc sinh hot ngay t u - iu m cc nc pht trin lm t hng chc nm qua. Tn dng kh sinh vt t phn gia sc v cht thi nng nghip Cng ti Farmatic Biotech Energy, c xy dng xong nh my phn hy k kh qui m ln u tin th trn Holsworthy thuc vng Bevon, Anh. y l mt d n do cng ti Farmatic, cng ng a phng
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v cc nng dn y hn vn xy dng. Nh my c xy dng t thng 2/2001 v theo k hoch, nh my c a vo sn xut vi cng sut y vo thng 6/2002. Nh my kh sinh vt Holsworthy c cng sut x l cao nht l 146.000 tn phn tru b, gia cm, k c cht thi lng thc hu c. Phn gia sc c thu gom t 25-30 trang tri a phng, cch nh my 10 dm. Cht thi lng thc sau khi ch bin c thu gom t cc c s ch bin lng thc vng Ty Nam. Jorgen Fink, Gim c iu hnh ca cng ti Farmatic tin rng, Anh c tim nng xy dng t nht 100 nh my. Cc nh my ny c th ng phn hy phn gia sc vi cht thi lng thc (nh Holsworthy) hoc c th ch phn hy cht thi lng thc v sinh hot. Cht thi lng Enretech-1 c 2 cng dng l cht thm du v ng thi phn hy sinh hc du. Sn phm c cha cc loi vi sinh tn ti sn c trong t nhin, khng b ci bin v gene. Khi c ngun thc n l cc hydrocarbon v m thch hp, cc vi sinh ny s pht trin nhanh chng v lng v "n" du, chuyn ha cc cht c hi thnh v hi. Vi sinh ch tn ti v pht trin trong x bng ca Enretech-1, khng th nui cy pht trin mi trng ngoi "ch" ca chng. Sn phm c sn xut t ngun nguyn liu tn dng li trong cng nghip ch bin bng. c tnh * Hp th nhanh cc hp cht hydrocarbon mi dng nguyn, nh tng tng phn hay b phn tn. Kh nng hp th gp 2-6 ln trng lng bn thn. * C lp cc cht lng m n hp th, khng nh li mi trng, do khng pht sinh ngun nhim th hai. * Phn hy hydrocarbon bng vi sinh t nhin c sn trong cc x bng ca Enretech-1. * Khng c hi i vi sc khe con ngi, ng thc vt v mi trng. * Hn hp Enretech-1 & du b hp th l cht thi thng thng, c th chn lp nh cht thi khng nguy hi do t cc tiu chun an ton ca b Mi trng M (USA EPA TCLP 1311, 9095A & 9096). * n gin v an ton khi s dng, khng cn chuyn gia hay hun luyn c bit. Phm vi s dng: Enretech-1 c s dng cho ng cu khn cp cc s c trn du trn t, x l ti ch t ct b nhim du.
187 Trng Vn Lung
Khi vic thu gom du trn bng cc bin php c hc (phao quy, bm ht, tm thm...) khng th thc hin c trn mt t, b sng, b bin, cc di ... b nhim du th Enretech-1 l gii php x l hiu qu kinh t nht v trit nht. Cc x bng ca Enretech-1 s hp th hydrocarbon ngay khi tip xc. Kh nng kt bao rt mnh l c tnh u vit gip c nh du trong cc x bng, loi tr nguy c du lan rng hay ngm su xung t, nh tng trong nc hay pht tn vo khng kh. Qu trnh phn hy sinh hc du ( b c lp) bi vi sinh Enretech din ra ngay sau . 70 - 80% lng du hp th b phn hy sau 2 thng. Trong iu kin thch hp, 80% hydrocarbon b phn hy sau 30 ngy. Vi sinh Enretech pht trin tt nht khi t nhim du iu kin nhit 25 - 300C, m 40%, pH 6 - 8. Khi nhit di 150C hay trn 400C, vi sinh ngng hot ng v pht trin. Thi gian hydrocarbon b phn hy hon ton nhanh hn rt nhiu so vi thi gian x bng Enretech t phn hy nn khng gy nguy hi cho mi trng. H thng x l nc thi trong cc b aeroten Cc b aeroten cn gi l phng php hiu kh, sc kh hay khng kh sinh hc. i vi phng php ny, vi sinh vt sinh trng trng thi huyn ph. Qu trnh lm sch aeroten din ra theo mc dng chy qua ca hn hp nc thi v bn hot tnh c sc kh. Vic sc kh y m bo cc yu cu ca qu trnh: lm nc c bo ha oxygen v duy tr bn hot tnh trng thi l lng. Nc thi ban u c tch s b cn, rc trc khi vo b iu ha, sau c bm t ng ln b aeroten cao ti. y cht hu c c phn hy nh vi sinh vt hiu kh c hot tnh cao v b phn phi kh hiu sut cao. Bn hot tnh c tch t b lng lin hp vi b phn ng ( gim chi ph xy dng v mt bng) mt phn c tun hon cn phn ln c x l b tiu hy bn. Nc thi sau x l t tiu chun thi c vo ngun tip nhn. Lm sch nc H Ty bng cy thy sinh Sau khi d n thay nc H Ty ph sn, thnh ph H Ni chuyn hng sang phng php sinh hc, t ri ro v r tin hn: trng cy thy sinh trong h. D n va c cng ti u t khai thc H Ty xut, d kin thc hin trong 24 thng (2004 - 2005) vi tng chi ph gn 5,4 t ng. Cc loi cy em trng s c la chn theo tiu ch sau: u tin v quan trng nht, phi chn nhng loi cy pht trin tt trong nc, di y h, trn mt nc, nm trong qun th thc vt vng chu th
188 Trng Vn Lung
Sng Hng c kh nng lm gim nhim mi trng. Th hai, to ra c cnh quan p. Th ba, c gi tr v kinh t. Sau khi trng, cng ti s chm sc, bo v, theo di, kim sot s sinh trng v pht trin, thu hoch sn phm, bn sn phm, nghim thu, tng kt d n. Cc loi cy thy sinh c trng gm: sen hoa cc mu, hoa sng cc mu (25ha), rau mung b, rong ui ch, rong tc tin, rong rp (5 ha cho cc loi rong, trng di y h). D n nhn mnh vic khng ch s pht trin trn lan, n mc ging nh nhng cuc xm lng, nh ca bo ty trong mt thi k, ca cc loi cy thy sinh ny, trnh tc ng ngc. Din tch ti a c php cho trng thy sinh - v phi thc hin c bng nhng tc ng mnh ca con ngi - l khng qu 25 ha, tc l 4,75% mt nc H Ty. Mt cuc hi tho c t chc ngy (21/11/2003), ly kin ca cc nh khoa hc u ngnh v d n. Theo GS. Mai nh Yn: "Nn lp mt vn cy thy sinh trn H Ty". Phng vin VietNamNet c bui tr chuyn vi GS. Mai nh Yn sau bui hi tho v nhng iu cn bn k hn xung quanh d n trng cy thy sinh, cng nh sng kin ca ng v mt vn cy thy sinh lu gi ngun gene thc vt cho H Ty v c vng ng bng Bc B. Dng bo lc sch nc h Xun Hng Trc tnh trng nc h Xun Hng b nhim nghim trng do cc ngun nc thi trn lu vc v, Ban Qun l v Khai thc Cng trnh thy li Lt th bo ng lot vi s lng ln xung cc h lng nm pha trn, ni cha ngun nc trc tip vo h Xun Hng. l cc h i C (phng 2), Cu St (Trng Trnh, phng 9), Hng Lc (Phm Hng Thi, phng 10). T my nm qua, ngun bo cc h ny c cho vt sch v iu gp phn gy mt cn bng sinh thi mi trng nc h. X l nc thi ca vt nui bng cc cy thy sinh Kh nng thch ng: Nc thi t cc tri chn nui cha khi lng ln cc nitrogen, phosphore v nhng hp cht v c c th ha tan c. Tht kh tch nhng cht ny khi nc bng qut tc hay lc thng thng. Ta c th x l chng mt cch hiu qu bng s dng cc loi cy va t chi ph li va khng nh hng mi trng. Hai loi cy hu hiu x l nc thi l bo lc bnh (water hyacinth) v c mui nc (water dropwort). Thi gian duy tr trong nc (Hydraulic Retention Time- HRT) c tc ng nht ca nc thi l khong 10 ngy trong ao h hay mt nc thong trng mt trong nhng loi cy thy sinh ny. Gii thiu v cy: C mui nc (water hyacinth - Oenanthe stolonifera)
189 Trng Vn Lung
C mui nc l loi cy leo lu nm, cn gi l cy cn ty nc (water celery). Loi bn a ca vng ng Nam , thn v l ca n c th n sng hoc chn nh mt loi rau. N sinh sn theo cch phn chia r v sinh trng tt nht trong mi trng nc nng cho ti su 20cm, hoc cc b ao v sui. Bo lc bnh hay cn gi bo Nht Bn, water hyacinth,(Eichhorma crassipes) Bo lc bnh c ngun gc Nam M, sinh trng nhanh v ni trn mt nc. Hoa mu tm c coi l cy trang tr mt s nc chu v sau tr thnh mt loi c di thy sinh chnh. N c th ti sinh rt khe v nhanh. X l nc thi: Nc thi ca vt nui cho chy vo b lng, cht thi rn lng xung y. Sau mt vi ngy cho phn nc trong chy vo b m c bo lc bnh hoc cy c mui nc. Mt nc trong b ny c cy che ph (mt t xp x 400cy/b). Nu l bo lc bnh, th b c th lm su tu . Cn loi c mui nc th nc nng mt cht, nn phi hn ch su ca b x l khong 30cm. C mui cn thi tit mt m cn bo lc bnh li thch thi tit m p. Cc kch c ca b ty thuc vo lng nc thi cn c x l. Chng hn, cht thi ca 10 con gia sc s khong 456lt. B s phi l 6m mi cnh v su na mt. Bo tm (Lemma japonica) v bo Nht Bn (Eichhornia crassipes) x l nc thi sinh hot, nc thi cc l m ng vt, nc thi ca l bn (theo Trng Vn Lung nm 2000). Qua kt qu phn tch cho thy khi x l nc thi bng cc loi bo nhim bn ca nc thi th hin BOD5 ch mc 9 - 20mg/l (gim t 92 96%), COD l 20 - 37mg/l, nitrite, nitrate v phosphate gim r rt, c bit NH4+ bo hp th t 90 n 99%. Trong mi trng nc thi cht hu c c phn gii thnh cc cht v c l thc n tt cho bo. Ngoi ra ngi ta cn c th s dng mt s loi to lc nh: Chlorella pyrenoidosa, Chlorella vulgaris v cc tp on vi sinh vt c sn trong nc thi x l nc thi sinh hot c hiu qu. Nc thi To sinh hotCt lc sinh hcB toThu hoch Nc x l
Bn to X l nc thi cc lng ngh bng lau, sy Lau, sy l loi cy c th sng trong nhng iu kin thi tit khc nghit nht. H sinh vt xung quanh r ca chng v cng phong
190 Trng Vn Lung
ph, c th phn hy cht hu c v hp th kim loi nng trong nhiu loi nc thi khc nhau, nh cc loi nc thi lng ngh. Phng php dng lau, sy x l nc thi do Kathe Seidel ngi c a ra t nhng nm 60 ca th k XX. Khi nghin cu kh nng phn hy cc cht hu c ca cy ci, ng nhn thy im mnh ca phng php ny chnh l tc dng ng thi gia r, cy v cc vi sinh vt tp trung quanh r. Trong , loi cy c nhiu u im nht l lau, sy. Khng nh cc cy khc tip nhn oxygen khng kh qua khe h trong t v r, lau, sy c mt c cu chuyn oxygen bn trong t trn ngn cho ti tn r. Qu trnh ny cng din ra trong giai on tm ngng sinh trng ca cy. Nh vy, r v ton b cy lau, sy c th sng trong nhng iu kin thi tit khc nghit nht. Oxygen c r thi vo khu vc xung quanh v c vi sinh vt s dng cho qu trnh phn hy ha hc. c tnh, s lng vi khun trong t quanh r loi cy ny c th nhiu nh s vi khun trong cc b hiu kh k thut, ng thi phong ph hn v chng loi t 10 n 100 ln. Chnh v vy, cc cnh ng lau, sy c th x l c nhiu loi nc thi c cht c hi khc nhau v nng nhim ln. Hiu qu x l nc thi sinh hot (vi cc thng s nh ammonium, nitrate, phosphate, BOD5, COD, coliform) t t l phn hy 92 - 95%. Cn i vi nc thi cng nghip c cha kim loi th hiu qu x l COD, BOD5, chrome, ng, nhm, st, ch, km t 90 - 100%. Theo Nguyn Quang Minh, v Khoa hc Cng ngh, b Xy dng, nc ta hin c khong 1.450 lng ngh truyn thng, tp trung ch yu ng bng Bc B, vi cc ngh nh ch bin sn phm nng nghip (lm bn, min, nu ru, ch bin tht gia sc, gia cm); sn xut, ti ch giy, st, nha, ha cht; sn xut gm, mc, kim kh. Ti nhiu lng ngh, nc thi ang l nguy c ln gy nhim nc mt, lm pht sinh nhiu mm bnh nguy him. Nc thi khng c x l m x thng ra sng, h, knh, mng... hay t b hoang ca lng. Vit Nam l t nc nhit i, kh hu nng m, rt thch nghi cho s pht trin ca cc loi lau, sy. Mt khc cc lng, din tch t nng nghip b b hoang cng cn kh ln. Do vy, vic p dng phng php x l nc thi bng lau, sy s rt hiu qu. X l nc thi nui tm bng rong bin y l ti khoa hc do Phm Vn Huyn, phn vin Khoa hc Vt liu ti Nha Trang, lm ch, c Hi ng Khoa hc Cng ngh tnh Khnh Ha nh gi tt.
191 Trng Vn Lung
ti xut mt s m hnh x l nc thi bng cc loi rong bin i vi tng loi ao a t hiu qu nui tm cao nht nh: s dng rau cu cc i vi loi ao, a c y ct hoc ct pha bn; rau cu ch vng i vi ao, a y bn; rong sn i vi ao m, vnh gn bin. ti c ng dng thc t ti cc ao cha nc thi ti cc khu nui tm s ng B (Nha Trang), Ba Ngi (Cam Ranh), kt qu cho thy, khi nhng ni ny c x l bng cch trng cc loi rong bin th hm lng nhng yu t gy nhim mi trng trong nc thi nui tm u gim t 60 - 80%. Nghin cu x l nc thi nh my thuc da bng bin php lc sinh hc Hin cc c s thuc da hoc khng c trang b hoc c trang b mt h thng x l nc thi nhng trong tnh trng khng hot ng v nhiu l do khc nhau. Do vy, nc thi ca cc n v ny u c x trc tip ra cc sng, h xung quanh c s sn xut, gy nhim trm trng. m bo cht lng ngun nc v sc khe cng ng, cc dng thi ny buc phi c x l trc khi x ra cc ngun tip nhn. Trong s cc gii php c th p dng x l nc thi ca nh my thuc da, phng php sinh hc l mt bin php kh thi do tnh tin ch, kinh t v an ton sinh thi. Bo co ny s trnh by mt s kt qu thm d k thut lc sinh hc x l nc thi loi ny. X l nc thi chn nui bng nm v vi khun C quan Hygefac Laboratories, Php, tch c 80 dng vi khun khc nhau m khi hn hp vo cht thi chn nui nga, gia cm, gia sc c th kh mi hi v ci thin gi tr lm phn bn ca cht thi. Cc vi khun a kh cng sinh, c bn trn th trng vi nhn hiu "Azofac", khi a vo cht thi chn nui lng s tc ng n vi khun k kh, l vi khun bnh thng pht trin trong cht thi v pht thi cc kh c mi hi. Tim nng ca cng ngh ny c phng Th nghim Quc gia Php cng vi cc k thut vin ca vin NIOSH ca M khng nh. Theo phng Th nghim Quc gia Php, c th gim ti 80% lng ammoniac v 90% hydrosulphite. X l cht thi lng bng Azofac cng cho php, nh gim pht thi ammoniac, hp th c nng nitrogen ln hn, lm tng gi tr lm phn bn. Mi y, cc nh khoa hc bt u s dng nm t c kt qu tng t. Mt nhm cc nh nghin cu do Jean Villard thuc trng i hc Paris V lnh o, tch c bn loi nm si t nhin: Mucor, Rhizopus, Aspergillus v Fusarium, c kh nng "tiu ha" cht thi lng. Bn loi nm ny tc dng tng hp vi nhau. Theo Villard, li ch ca
192 Trng Vn Lung
vic s dng nm l chng c nhiu trong cc enzyme. Sau khi x l trong b phn hy sinh hc, lng sinh khi cn li t hn nhiu so vi x l bng vi khun. Cc th nghim sn xut th trang tri cho thy, phn sau khi c x l, hon ton khng c mi v gim c 80% cht nhim. Chi ph lp t thit b khong 4,5 USD cho 35 foot khi (khong 10 m3) x l, trong khi cc h thng x l phc tp hn c th t gp 3 ln. D n ny do T chc Euralis (t chc hp tc ca cc nh chn nui v nhn ging) ti tr. Qui trnh x l cht thi lng ca Euralis gm hai pha. Trc ht, cht thi lng c x l vi vi khun k kh v sau c x l vi nm trong mi trng a kh. Trong bn phn hy sinh hc u tin, vi khun chuyn ha nguyn liu c nitrogen ha trong cht thi lng thnh cc nitrite. Trong bn phn hy th hai, nm hp th nitrite, gii phng ra kh dng nitrogen trong kh quyn. Lu hunh, ngun gy mi nhiu th hai trong cht thi lng, c to thnh phc cht trong sinh khi cn li. Nm cng tiu th mt phn ln phosphore v kalium cha trong cht thi v thu gi cc kim loi nng, nht l ch, thy ngn v cadimium. Vic kh c COD, nitrogen, phosphore v kalium to ra kh nng s dng cht thi x l lm nc ti. X l nhim mi trng khng kh S dng mt cch ng n thm thc vt, cy g, cy bi vi mc ch phng chng nhim mi trng khng kh l bin php quan trng trong iu kin hin nay. Quan trng nht l xy dng mt cch c h thng thnh ph v cc khu cng nghip, nhng khu rng, khu cng vin p ng nhng yu cu v kin trc qui hoch. Vn x l nhim mi trng khng kh hu nh cha c cng trnh no c sc. Ngi ta ch trng vic trng cy xanh cc thnh ph va to cnh quan mi trng, va hn ch c kh CO2. Ngay c M, quc gia thi rt nhiu kh thi cng ch c gii php tm thi l chn CO 2 di y i dng. Qua kt qu nghin cu, c th ni rng vn x l cc loi cht thi l rt cp bch v cn thit. Vic ng dng cng ngh sinh hc trong x l l bc i ng cho vn mi trng ngy nay. thc hin tt cng ngh ny, chng ti mnh dn a ra mt s kt lun v ngh sau: p dng cc phng php sinh hc l cng ngh c tnh kh thi cao, c nhiu u im v an ton v sinh thi. x l c hiu qu cn lng ghp, kt hp nhiu phng php x l khc nhau. Lm th no va tit kim chi ph li va c hiu qu trong bo v mi trng.
193 Trng Vn Lung
Cn c s lin kt gia cc nh khoa hc v cc doanh nghip, cn thc y hn na nhng ng dng em li li ch kinh t. Cc nh nghin cu cn u t nhiu hn na v nhng ti c trin vng lu di, c kh nng ng dng vo cng tc x l mi trng v bo v mi sinh. TI LIU THAM KHO Hong Hu, 1996. X l nc thi. Nxb Xy dng. H Ni. B Hng, 2003. X l nc thi nui tm bng rong bin. Bo Nhn dn, ngy 26/5. Phm Th Ngc Lan, Phm Th Ha, L Kim Bng, 1999. Tuyn chn mt s ging x khun c kh nng phn gii cellulose t mn rc. Bo co khoa hc Hi ngh Cng ngh Sinh hc ton quc ln th nht thng 12/1999, Nxb Khoa hc v K thut H Ni, tr 177-181. Trng Vn Lung, 2000. Nui bo Nht Bn (Eichornia crassipes), bo tm (Lemma japonia Landolt) v bo ci (Pistia stratioles L.) x l nc thi Tha Thin Hu. Tp ch Khoa hc, i hc Hu, S 10. tr: 103-107. Trng Vn Lung, Nguyn Ngc Thanh, 2003. Thm d mt s bin php sinh hc x l nc thi t qu trnh sn xut ca nh my ch bin tinh bt sn Qung Nam. Bo co khoa hc Hi ngh Cng ngh Sinh hc ton quc ln th 2, H Ni thng 12/2003, Nxb Khoa hc v K thut H Ni, tr 3 1 3 - 3 1 6 . Trn Vn Nhn, Ng Th Nga, 1999. Gio trnh cng ngh x l nc thi. Nxb Khoa hc K thut. H Ni. ng Xuyn Nh, Nguyn Ph Cng, Dng Hng Dinh, 1999. ng dng to v ct lc sinh hc trong x l nc thi snh hot qui m nh. Bo co khoa hc Hi ngh Cng ngh Sinh hc ton quc ln th nht thng 12/1999, Nxb Khoa hc v K thut H Ni, tr: 192-197. Nguyn Quc Thng, ng nh Kim v cng s, 2003. Nghin cu kh nng hp th kim loi nng Cr v Ni ca bo ci (Pistia stratiotes L.) t nc thi, Nhng vn c bn trong sinh hc, Bo co khoa hc hi ngh ton quc ln th 2, Nxb Khoa hc v K thut H Ni, trang 401 - 404. Trn Vn Ta, Nguyn Tin C, ng nh Kim, H T Cng, Hong Th Bo, 2003. Nghin cu x l nc thi ch bin thy sn bng phng php k kh. Bo co khoa hc Hi ngh Cng ngh Sinh
1. 2. 3.
4.
5.
6. 7.
8.
9.
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hc ton quc ln th 2, H Ni thng 12/2003, Nxb Khoa hc v K thut H Ni, tr 141-143. 10. Trn Cm Vn, Bch Phng Lan, 1995. Cng ngh vi sinh v bo v mi trung. Nxb Khoa hc & K thut. Trung tm Giao lu quc t v Vn ha Gio dc v Khoa hc (CCES), H Ni. 11. Ch phm BRF-1 AQUAKIT v BRF-2 AQUAKIT x l nhim du,
www.oilspill.com.vn 12. Dng bo lc nc H Xun Hng, www.vnexpress.net 13. Hoa ng sc chng nhim ch trong t, www.vnn.vn 14. Kh mi rc bng ch phm sinh hc, www.hanoitv.org.vn 15. Nghin cu x l nc thi nh my thuc da bng thp lc sinh hc, www.biotechvn.com.vn 16. Seraphin-Cng ngh x l rc thi sinh hot Vit Nam, www.vnexpress.net 17. X l cht thi chn nui bng nm v vi khun, www.vista.gov.vn 18. X l nc thi lng ngh bng lau, sy, www.vnexpress.net 19. X l rc thi bng giun, www.vnexpress.net
195 Trng Vn Lung
Chng X
Cng ngh sinh hc trong vic bo tn v pht trin ngun gene qu him
1. Lu gi ngun gene
ng lc ch yu ca cuc cch mng CNSH, cn gi l cuc cch mng xanh ln th hai li l cc t chc t nhn mc d hu ht cc nghin cu c bn u l c tin hnh cc trng i hc Quc lp v cc trm Nng Lm nghip. Php vic phn phi thng mi ging cy nhit i thng qua k thut vi nhn ging phn ln do cc c quan cng cng nh trung tm Hp tc Quc t v Nghin cu Nng hc cho Pht trin (CIRAD) v cc chi nhnh Vitropic v Tropiclons ca trung tm kim sot. T nhn ha cc kt qu nghin cu CNSH c ngha l cc kt qu nghin cu ny khng cn l mt b phn cu thnh ca tri thc khoa hc k thut, mt ti sn chung cho mi ngi na. Qu trnh t nhn ha nh vy c lin quan n ngun gene v ti nguyn di truyn thc vt rt cn cho cng tc lai to v nui cy m thc vt. Cc dng lai c c tnh mong mun v c dng vo ti to ging mi. Cc ging thng mi mi hoc cc dng t bo, cc sn phm mi v cc hng s dng mi ca cc sn phm ny. Cc qui trnh k thut mi c lin quan n vic to ra ging hoc sn phm mi. T nhn ha lm xu i kh nng tip cn thng tin v b quyt k thut cng nh kh nng tip nhn cc ging cy trng, d dng sng hay gi ng lnh, hn ch mc phong ph ca i mi cng ngh. Gia nhng nm 1980, c 14 cng ti v tp on ging ln chim khong 20% th trng, cc nc kinh t th trng.. Doanh thu hng nm ca h hng triu USD, sn xut, lu gi v bn ht ging. Cc cng ti ny huy ng vn khng l (t 18 n 30 t USD) trn th trng th gii. lm c s cho vic chn ging cy trng, cho n nay cc nc tin tin c cc tp on ging chuyn gi ging cung cp cho cc nh chn ging lm vt liu ban u cho vic lai to. Cc tp on ging ln nht th gii l tp on ging la vin Nghin cu la Quc t Philippines c 120.000 ging , tp on ging la m vin Cy trng Lin X (c) c trn 20.000 ging, tp on ging ng trung tm Quc
196 Trng Vn Lung
t Ci bin ng v la m Mexico c hn 10.000 ging, tp on ging cao lng vin Khoa hc Nng nghip n c hn 8.000 ging. v.v. Thng mi ging cy trong tp on ch c vi c tnh tt. Do vy, mun c mt ging cy trng l tng cha ng cc gene tt ca nhiu ging phi tn thi gian di mi lm c. Do vy, trong cng vic ny phi lm dn do nhiu th h cc nh chn ging ni tip cng vic ca nhau. Vic tng hp cc gene mang c tnh tt ca cy trng thng c lm bng phng php lai v phi lai trn nhiu cp lai phi hp vi nhau tng i mt mi mong t c kt qu tt. Sau ny, cc nh khoa hc pht hin rng b v m c c tnh tt th con lai cng li ci tt ca b v m. l tc dng cng ca gene, v nu con lai tha hng cc c tnh ca nhiu b v nhiu m th con lai cng tt hn - gi l lai tng hp. V vy cho nn vic lu tr ging lu tr ngun gene sinh hc l iu ht sc cn thit cho ngy nay v cho c th h mai sau. V vy, l gi ngun gene sinh hc ex-situ v in situ (bn ngoi v ti ch) l iu ht sc lu i vi cc nh sinh hc, c bit l cc nh di truyn hc.
2. Bo tn gene v Ngn hng gene

Trc ht cn xc nh ngun gene l g? Theo Qui ch qun l v bo tn ngun gene ng vt, thc vt v vi sinh vt c b Khoa hc Cng ngh v Mi trng ban bn ngy 30 thng 12 nm 1997 th ngun gene l nhng sinh vt sng hon chnh hay b phn ca chng mang thng tin di truyn sinh hc, c kh nng to ra hay tham gia to ra ging mi ca thc vt, ng vt v vi sinh vt. T nh ngha ny c th thy r bo tn ngun gene chnh l bo tn cc vt th mang thng tin di truyn nhng vt liu ban u c kh nng to ra ging mi. iu quan trng khi bt tay vo bo tn ngun gene l phi xc nh c mc tiu bo tn. Mc tiu khc nhau th phng php bo tn v i tng bo tn cng khc nhau. Cho n nay, mc tiu bo tn trong nng nghip bao gi cng xc nh l s dng cho cng tc chn ging v gy ging trc mt v trong tng lai. V vy, vic bo tn ngun gene bao gi cng c tp trung gii quyt cho cc loi cy trng ch yu m khng bo tn cho cc loi cy c khng c ngha kinh t. u v bao gi bo tn gene cng nhm s dng cho cng tc ging trc mt v lu di c hiu qu hn. V vy bo tn ngun gene bao gm vic bo tn cc sinh vt nguyn vn ln nhng b phn c dng cho lai ging v nhn ging.
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Nhng nm va qua, nhn dn ta do thiu lng thc phc v cho cuc sng nn ch n nng sut cy trng vt nui, cha thc s ch n phm cht. Chng ta cng cha c ni lu gi ging, do ngn hng gene ca chng ta cn rt ngho nn. Ngn hng gene l dng k thut phc ch li DNA ly ton b DNA trn mt loi t bo sinh vt hoc ton b DNA trn nhim sc th cy ln gene vn chuyn sau chuyn dch n t bo c tr tin hnh bi dc thnh h v tnh. Sau lc thnh lp ngn hng gene c th cho tin hnh sinh sn v tnh lm cho ton b gene nhp vo t sinh sn v tnh, chnh l ngn hng gene. Khi c ngn hng gene, h cn n gene no hoc on no c th ct ghp hoc phc ch, chnh v vy m khi thc hin s d dng hn. Nu gene gc, lc cn bt buc phi tch n ra, vic tch cng khng phi d dng. Nu chng ta thnh lp sn ngn hng gene ca cc sinh vt khc nhau th cng vic tin hnh s thun li. T nm 1974, cc nh khoa hc thnh lp ngn hng gene ca trc khun i trng, men gim, rui gim, g, th, u tng. S lng gene cha trong ngn hng gene v ln b ca t hp gene c lin quan vi di ca on DNA ca h sinh sn v tnh. Sau khi thnh lp ngn hng gene th vn quan trng l lm th no gi gn n. bo qun tt ngi ta thng dng phng php sinh sn ca vi khun lm cho cc vi khun cha nhiu DNA khng ngng tng ln. Do sc sinh sn ca vi khun ca cc loi khc nhau th ging nhau, loi sinh si ny n chm th phc ch t. Lc u ch c mt vi khun cha DNA nh sn, sau thi gian nui cy, s lng vi khun tng ln n hng trm triu, nh th s lng DNA cng tng ln hng trm triu. Chng ta ch cn ra, tch vi khun trong mi trng nui cy, t ni nhit thp c th bo qun lu di. Khi cn s dng ch cn tch vi khun ra l c. Ngn hng gene ca thc vt bc cao v ng vt bc cao dng ct gene duy nht l enzyme cng c. chuyn ha ca enzyme rt cao, mi loi enzyme mang tnh c hiu ring cho mt gc base trong nucleotide m khng th dng chung cho 1 loi gc base khc, tng t nh chia kha v kha. Do enzyme ct c c tnh ny nn n tr thnh cng c chuyn dng ct gene. Cho d DNA di bao nhiu ta cng c th dng enzyme ct ly on DNA m ta cn. Nh th gene c ct, vy lm th no cc gene lin kt li vi nhau, ta s dng enzyme lin kt tng t nh keo dn ni 2 gene li vi nhau.
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Sau khi 1 chui DNA di b ct, vt ct c dch dnh gi l on cui dch. Khi ni 2 on ca gene khc nhau li th chng khng th gn mt cch nhanh chng c. Phi dng enzyme lin kt mi gn 2 on ca gene khc nhau li, to thnh chui chc chn. Nh th, mt phn t DNA ti to c ra i. Cc phng php bo tn v pht trin ngun gene thc vt v ng vt Vit Nam l mt nc nhit i m, gi ma, thin nhin phong ph v a dng ngun li ng vt v thc vt. Chng tr thnh nhng ngun gene qu him v nhiu loi a vo Sch , c nguy c dit chng. Vic bo tn v pht trin ngun gene tr nn v cng cp bch i vi nc ta. Chng ta c th nu ra mt vi v d n gin chng minh cho tnh cht c o v di sn thin nhin ca dy t min Trung. l s xut hin ng c bo v ca cc loi cy bn a qu him nh kim giao (Podocarpus ananmensis), kin kin (Hopea pierrei Hance), g lau (Sindora tonkinensis A.Chev), ch en (Parashorea stellara Kurz), hunh (Tarrietia cochinchinensis Pierre), cm lai (Dalbergia bariaensia), trc (Dalbergia cochinchinensis), trm hng (Aquilaria crassna Pierre ex Lecomte),du Hasel cn gi l du ri (Dipterocarpus hasseltii Blume), i bay (Sterculia lychnophora Hance), re hng cn c tn khc d hng, x x (Cinamomum parthenoxylon (Jack) Meissn.), lim xanh (Erythrophloeum fordii Oli in Hook), l khi (Ardisia silvestris Pit.), by l mt hoa (Paris polyphylla (Smith) Raf.), vng ng (Coscinium fenestratum (Gaertn.) Colebr.), v.v. V ng vt th c: chim tr (Lophura edwardsi), g g (Arbolophila merlini), tr sao (Kheinardtia ocehlata), g li lam mu trng (Lophiera ediwardsi), g li lam mu en (Lophiera imperalis), voc ch v (Lygethrix nemanneus), voc H Tnh (Presbitis fracoisi hatinh), vn (Hylobates concolor gabrrella), ch si (Cuon alpinus), bo gu (Neofelis nebulosa), bo hoa mai (Panthera pardus), h (Panthera tigris), v.v. Nhng ngun gene qu him ny khng nhng cn thit phi gi gn bo v m cn cn nhn ln din rng nui trng trn cc vng i ni. Cc phng php pht trin ngun gene V thc vt: Nhn ging in vitro l mt trong 4 lnh vc ng dng chnh ca cng ngh t bo thc vt, l lm sch virus, nhn nhanh cc ging cy trng qu, sn xut v chuyn ha sinh hc cc hp cht t nhin ci tin v mt di truyn cc ging cy mang li hiu qu kinh t cao. K thut nhn nhanh c ng dng trong nhiu lnh vc:
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- Duy tr v nhn nhanh cc kiu gene qu him lm vt liu cho cng tc chn ging. - Nhn nhanh v duy tr cc c th u dng tt cung cp ht ging cc loi cy trng khc nhau nh cy lng thc c c, cc loi cy rau, hoa, cy cnh, cy dc liu thuc nhm thn tho. - Nhn nhanh cc kiu gene qu him ca ging cy lm nghip, gc ghp trong cy n qu, cy cnh. - Nhn nhanh iu kin v trng, cch li ti nhim kt hp vi lm sch virus. - Bo qun tp on nhn ging v tnh, cc loi cy giao phn trong ngn hng gene. Cc phng php nhn ging v tnh bng gim cnh, chit, ghp, tch cy con t c th m u l nhng k thut to ra dng v tnh. - Th phn v nui cy phi in vitro Tnh bt hp ca giao t c th vt qua c bng ni dung k thut th phn trong ng nghim: . Tch bu qu v non ra nui cy trong iu kin v trng. Ht phn c ly v t ln bu nhy. Th phn thnh cng, phi s sinh trng v pht trin trn mi trng nui cy. (Kch thch ht phn ny mm, kch thch sinh trng ca ht phn, nui non v th tinh non, nui hp t thnh ht) . iu kin c bn l nui cy thnh cng bu qu, non phn lp ch ng iu khin qu trnh ny mm ca ht phn trong mi trng nui cy v trng. - Ngoi ra ngi ta cn dng phng php to cy n bi in vitro, tch v nui cy t bo callus, bin np gene vo t bo callus, to t bin v chn dng t bo, s dng khi u crown gall v plasmid Ti, bin np thng qua Agrobacterum tumerfaciens, nhn ging v sn xut ht nhn to, to ging mi bng phng php chn dng soma trong nui cy m t bo, v.v. Bo qun in vitro ngun gene thc vt Mc ch c bn ca bo qun ngun ti nguyn di truyn thc vt l duy tr s phong ph a dng di truyn trong loi, gia cc loi trong h sinh thi ni chung lm c s cho vic khai thc s dng trong hin ti hoc trong tng lai. Da vo c tnh sinh sn cc loi cy trng c xp thnh: - Cy sinh sn hu tnh bo qun nhit m v m thp. - Cy sinh sn hu tnh mt sc ny mm bo qun nhit thp kh. - Cy nhn ging v tnh mt kh nng sinh sn hu tnh hon ton
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- Bo qun ti ch cho php cy trng sinh trng trong iu kin sinh thi t nhin l nhng cy sinh sn v tnh cy hoang di. - Cc c quan sinh sn ca cy trng c bo qun trong iu kin nhn to ti u: . Trn vn m . Trong kho lnh . Bo qun in vitro . Bo qun DNA Vic la chn cc phng php bo qun ph thuc vo bn cht cy trng. Bn cht cy trng quyt nh cc gii php bo qun :
Yu t sinh hc 1. Cy lu nm 2. Cc cy hng nm Nhng phng php bo qun thch hp Ngn hng gene ng rung, bo qun in vitro ht v ht phn Bo qun ht-ht phn
Ngn hng gene ng rung v in vitro bo qun ht 3. Ht orthodox Ngn hng gene in vitro, ti ch v trn ng rung 4. Ht recalantrant Ngn hng gene ng rung in vitro v bo qun ng kh 5. Cy nhn v tnh vi ht v sinh Ngn hng gene ng rung 6. Cy nhn v tnh vi ht c v tnh Bo qun ht-ht phn thng - Bo qun in vitro ng kh 7. Cy c ht phn vi sc sng cao Bo qun ht phn 8. Cy c iu kin nui cy m Tm cc phng php bo qun thay th 9. Cy mc n nh di truyn cao Tm phng php bo qun thch hp
Ngn hng gene in vitro bo qun thng s dng 2 phng php: - Nui cy bo qun ng kh. Gii php ny c pht trin trong vng 10-15 nm tr li y c bt u bng bo qun callus, ph thuc vo mc ch nghin cu sinh l ha sinh. Da vo nguyn tc , chng c chuyn sang nui cy bo qun chi mm phi v cy con hon chnh phc v cng tc nhn nhanh sn xut cy ging, xy dng ngun gene ca cy trng. - Bo qun in vitro xut hin ng thi vi vic s dng phng php truyn thng. Trong thi gian ti, bo qun in vitro s cho ra i cc ngn hng gene in vitro. Bo qun in vitro xy dng ngun gene thc hin 2 nhim v:
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. Bo qun cc tp on hot ng nhm cung cp vt liu phc v cng tc nghin cu v trao i ngun gene gia cc ngn hng gene. . Bo qun tp on c bn bo tn ngun gene lu di b sung vt liu cho cc tp on hot ng. V ng vt: Hin trng sinh sn v tnh ng vt trong t nhin khng ln ng vt v ngi. Chng ta thng gp nhng tp hp gm 2-3-4 ngi ging nhau. l hin tng song sinh cng trng, nm ch em Dionne sinh nm 1934 Canada hon ton ging nhau. Mt ngi cht v bnh ng kinh, 1 ngi i tu, 3 ngi ly chng v c con. Nh vy, chng l bn sao ca nhau nhng vn l con ngi khc nhau. xem xt 2 c th, ngi ta s dng phng php xt nghim DNA, ch cn ghp da cho nhau l c th bit c. Hin tng 1 phi tch lm nhiu phi v sau mi phi chnh thnh 1 c th c thy nhiu loi nh ong k sinh. Cng nh trong trng trt, chn nui, ging l khu quan trng hng u quyt nh sc sn xut, nng sut, cht lng ca vt nui. Nh vy, nng cao nng sut, phm cht ca vt nui cn ci to ging v cn c ging tt. C nhiu bin php ci to ging: phng php c in l chn lc, lai to ging to ra ging tt, phng php hin i l chuyn phi, chuyn gene. Nh vy, bo tn v pht trin ngun gene ng vt, ngi ta dng mt s phng php sau: - Cng ngh chuyn phi to ging mi bao gm th tinh trong ng nghim (ngy 29 thng 11 nm 2002, 2 con B con Vit Nam ln u tin ra i trong ng nghim do cc nh khoa hc vin Cng ngh Sinh hc H Ni cho th tinh ng nghim sau chuyn phi vo b ci lai Sind) , th tinh nhn to, chuyn phi (x l con ci cho phi, gy hin tng siu non: cho rng nhiu trng, k thut thao tc phi, k thut x l vi con ci nhn phi). - Cng ngh gene to ging mi (thay i b gene ca t bo ng vt bng cch gn DNA ngoi lai vo non bo c th tinh, b gene ca phi s thay i do on DNA ngoi lai ha nhp vo b gene sn c ca phi. bo tn v pht trin ngun gene ng vt, ngoi k thut phi, k thut gene, ngi ta xy dng c mt s qui trnh k thut khc to ging mi. Thng 2 nm 1997, Ian Wylmut v cc ng nghip ti vin Roslin, Scotland thnh cng trong vic nhn bn v tnh t 1 t bo tuyn v. T 1 t bo ca con cu ci c tn l Finn Dorset tin hnh nui cy
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trong phng th nghim. Ly trng ca con cu ci khc ht nhn ch cn li t bo cht sau cho kt hp t bo tuyn v vi trng khng nhn. K thut thut ny gi l truyn nhn t bo c th (somatic cell nuclear transfer). Sau 6 ngy cy hp t vo t cung con cu ci khc mang h thai. Con cu ny sinh ra cu Dolly ( ly theo tn din vin Dolly Parton) mang c tnh ging cu ci m. Nh vy, t 1 t bo trng thnh to ra bo thai ri to ra c th mi.. C ngha l t 1 t bo c th ti bn li ton b c th nh k thut nhn bn gene. S MINH HA K THUT SINH SN V TNH NG VT
Loi nhn kh t bo trng T bo sinh dng v t bo trng loi nhn
Phng php thng s dng kch thch in Phng php thng s dng kch thch in
Phng php thng s dng kch thch in
Cu Dolly sinh ra bng sinh sn v tnh
Cu con c bn gene ging ht m sinh nm 1997
Nui cy trong ng nghim to thnh phi nang. Cy chuyn phi
Dung hp t bo bng shock in
Hnh X. Sinh sn v tnh ng vt Nh vy, cng nh thc vt, t bo ng vt cng mang tnh ton nng. ng vt cng c th lai v tnh, sinh sn v tnh, tuy nhin c phc tp hn so vi t bo thc vt. Cc nha khoa hc cn tm cc bin php mi tng nng sut vt nui. Thng 2 nm 1994, ti M mt loi hormone tng trng ti t hp ra i. Ti trng i hc Cambrige (Anh) thnh cng trong vic to ra b c, ci theo mun. Va qua ti vin Cng ngh Sinh hc H Ni cng to ra nhng con b c b ci theo mun.
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Cc nh khoa hc Cambrige to ra trong c th con ln c gene ngi nhm mc ch ly ph tng ca chng cy ghp thay th cc b phn h hng ca con ngi. Ngy 24/11/1999, ch mo Jazz ging hoang d chu Phi cho i ti New Orleans (M), mo m Cayenne li rt ging mo nh. y l kt qu nhn ging th qu him ca trung tm Nghin cu cc loi b e da Audubon-New Orleans. Cc nh khoa hc ly tinh dch v trng ca mo c v mo ci ging hoang d chu Phi ang c nguy c tuyt chng, sau cho th tinh trong ng nghim. u tin, phi thai th tinh trong ng nghim c nui 5 ngy trong lng , sau c bo qun 1 tun -196oC. K tip cc nh khoa hc gii ng phi thai ri cy 8 phi vo t cung con Mo ci nh Cayenne, 70 ngy sau Mo Jazz cho i. Nm 2000, trn th gii cn cha ti 1.000 con gu trc ln, trong c khong 100 con ang sng trong vn th. Mt gu trc ci va cht. Cc nh khoa hc Trung Quc ct ly mt mu da ca n v ngm vo azote lng -196oC. (Theo mt s kin cho rng cch tt nht l ng lnh tinh dch con c v trng con ci mu vt lu gi c ton b h di truyn). Hin nay mt s loi gia sc nh cu, b, d, ln v loi chut nhn bn v tnh. Nu chng ta bo qun mu vt th c nguy c tuyt chng, bit u, mt ngy no c th cu ly ging ni s th ny. tng ca cc nh khoa hc Trung Quc ang ph bin nhiu ni trn th gii vi nhiu cch lm khc ch khng hn ch l nhn bn v tnh. V d mt gii php mi bo tn ng vt qu him S T vng ni Atlas, Bc Phi, ch si Tasmanta (chu c), hi cu trn qun o Antilles, nga vn u, d rng ni Pyrenees, Ra qun o Seychelles (n Dng), u tuyt chng. Chim cu li trn o Mauritius c pht hin nm 1658. Ch trong vng 80 nm loi chim c thn hnh bo trng trc ln lt h cnh xung bn n ca thy th thng qua li n Dng.Trong 500 nm qua, hn 720 loi ng vt tuyt chng do con ngi sn bt ly tht, ly lng hoc ph rng. T hn mt th k nay, khng cn con nga vn u no sng st trn tri t. Mt d n khi phc ging nga vn u qu ny sp c kt qu. Cng lc d n thnh lp cc khu bo tn Ha Bnh chu Phi cng ang ro rit c thc hin n tip du khch. Nga vn u c thn hnh mu ht d, khng to vn en trng ton thn m ch c cht t vn mu hi trng u v c. Chng sng Karoo (Nam Phi). Do him c nhn dn bn a ra sc tiu dit loi th hoang d ny by gia sc ca h c c n. Chng b tuyt chng. Hin nay ch cn 20 hin vt nga u vn nhi rm c bo qun trong
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mt s bo tng trn th gii. Reinhold Rau (chuyn gia nhi da ng vt bo tng Nam Phi) cht nh ra trong sch ca nh ng vt hc ngi c Lutz Heck c vit: khi nga vn ng bng c chn lc lai vi nhau s cho ra ng vt ging nh nga vn u (NV) tuyt chng. Reinhold Rau cho rng NV khng phi mt loi ring bit m ch l phn loi ca nga vn ng bng. Khi 2 loi khc nhau lai vi nhau th h i con s v sinh. Ngc li hai phn loi khc nhau trong cng mt loi lai vi nhau s cho ra th h lai mang gene ca b m. Reinhold Rau suy lun: ging nga vn ng bng ang sng Karoo s mang t nhiu gene gc ca NV v ch cn cho lai kho lo s tp trung c y b gene c trng ca NV, t vic khi phc ging nga ny ch l trong tm tay. NV v nga vn tng cn vi nhau c nhiu c tnh trung gian trn b lng. T nhng suy on c nhiu ngi ng h v d n nga vn u ra i. T phn tch mu DNA ca NV, Nga khng vn, nga vn ni, nga vn ng bng thy rng NV l phn loi ca nga vn ng bng nh Reinhold Rau d on. Cc nh khoa hc tin hnh tuyn chn v cho lai to t nm 1987. Hin by Nga th nghim ang sng 7 ni ti Nam Phi. Nh sinh hc Eric Harley thuc i hc Cap (Nam Phi) c tham gia d n NV nhn nh: Kt qu s t c khi th h nga th t ra i vo nm 2003 v phi mt 30 nm na chn gene c trng ca nga vn u. Thng 7 nm 2004, ln u tin trn th gii, Anh thnh lp ngn hng gene ca nhng ng vt hoang d. T c s ny ngi ta khng nhng c kh nng lu gi nhng ng vt c nguy c tuyt chng hin nay m cn c th nhn bn li nhng ng vt tuyt chng, k c nhng ng vt mt i t thi xa xa. TI LIU THAM KHO 1. Nguyn Mng Hng. 2004. Cng ngh t bo phi ng vt. Nxb i hc Quc gia H Ni, H Ni. 2. ng Hu Lanh. Trn nh Min. Trn nh Trng. 1999. Cng ngh sinh hc vi cng tc ging vt nui, b. Trong: C s di truyn chn ging ng vt (ng Hu Lanh, ch bin). Nxb Gio dc H Ni, tr: 433-469 3. L nh Lng, Quyn nh Thi. 2003. K thut di truyn v ng dng. Nxb i hc Quc gia H Ni, H Ni.
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4. 5. 6. 7. 8.
Sasson Albert, 1988. Biotechnologies and development Cng ngh sinh hc v pht trin. Ngi dch: Nguyn Hu Thc, Nguyn Ln Dng v mt s dch gi khc. Nxb Khoa hc & K thut H Ni. Bernard R.Glick, Jackj Pasternak. 1994. Molecular BiotechnologyPrinciples and Application of Recombinant DNA. ASM Press. Washington D.C. Chopra V.L, Anwan Nasin. 1990. Genetic Engineering and Biotechnology, Oxford and IBH Publishing CO.PVT, Ltd. Dalton. D.C. 1982. An Introduction to Practical Animal Breeding. Granada Publishing Limited, Great Britain. Peregrino G. Duran. 2002. Technical aspects of the recovery, handling and transfer of embryos. Copyright FFTC1970-2002. (http://www.fftc.agnet.org/library/article/tb151a.html).
CHSH vi s pht trin bn vng
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5
CNSH VI S PHT TRIN BN VNG Chng XI
Cng ngh sinh hc vi vic hp tc quc t v khu vc

1. Tc ng ca tin b CNSH i vi cc nc ang pht trin
Nhiu nc ang pht trin c c mt ti sn cng ngh v kinh nghim pht trin cng ngh sinh hc, song nhiu nc khng to c th mnh ca mnh bi s ln mnh vt bc ca cc nc cng nghip pht trin. C l cc ngnh nng nghip v cng nghip thc phm chu nh hng nhiu nht bi vic thng mi ha cc sn phm cng ngh sinh hc cnh tranh vi mt hng thc phm do cc nc ang pht trin sn xut. Xu hng tp trung ha trong sn xut v thng mi ha ging mt s cy ng cc c th hin kh r. Hin c khong 20 cng ti a quc gia (ch yu l cng ti Ha du v Dc phm) nm gn ht c phn ca cc cng ti sn xut ht ging, cung cp phn bn, ch phm bo v thc vt v thuc th y. Li nhun ca vic kinh doanh cc sn phm ny gia tng nhanh v chnh cc cng ti ny l ngi hng dn phn ln. Ngoi vic kim sot cc cng ti sn xut ging, cc cng ti a quc gia u t rt mnh vo vic nghin cu nhng vn v CNSH mi nhn p dng cho cy trng. Do vy, cng c th ni cc cng ti a quc gia ni trn chnh l nhng ngi tin phong trong cuc cch mng CNSH. Hin cc nc ang pht trin ang c xu th s dng t ging mi. Tuy nhin, ngoi hiu qu khng th chi ci v kinh t th vic lm dng ging mi mt cch thi qu v khng c nh hng ng n s lm cho ngun ti nguyn di truyn ni chung ca cc nc ang pht trin b suy gim. Bn cnh , tc ng ca vic sn xut cc cht t nhin hu ch bng sn phm CNSH cng c nh hng rt ln i vi cc nc ang pht trin. V trc y, cc nc cng nghip vn phi nhp nguyn liu thc vt cha cht th cp t cc nc ang pht trin. Tuy nhin, trong thi gian gn y, cc cng ti Ha cht v Dc phm cc nc pht
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trin (c bit l Nht Bn, c v M) bt u s dng k thut nui cy m v t bo thc vt sn xut qui m ln cc cht trao i th cp, v d cht gim au codeine, thuc tr su pyrethrin, tinh du Hoa nhi, vinblastin v vincristin t cy Da cn Catharauthus roseus cha bnh mu trng, cht b ginsenoside trong r cy Nhn sm,Vn ny s e da nghim trng cc phng php trng v thu hi cc hp cht cc nc ang pht trin. Cng bng cc phng php CNSH sn xut cc cht ngt khng phi ng, v hin trn th gii ang thnh hnh xu hng tiu th cc cht ngt khng phi ng cha t calo lm cho gi ng gim lin tc t nm 1985 n nay. Ngoi ra, ngi ta cn bit kh nhiu cht ngt t calo khc ang hon thin cng ngh sn xut chng, c bit l cng ngh sn xut sirofructose t tinh bt. Trong tng lai gn, sirofructose s thng tr th trng cht ngt th gii v nh vy s nh hng tiu cc i vi cc nc ang pht trin xut khu ng truyn thng. Hin nay kh nhiu loi thc phm vn ch yu c sn xut bng cng ngh truyn thng, nhng trong tng lai gn chng c sn xut bng cng ngh mi. V d: hin nay cc chuyn gia CNSH ca hng Nestle (Thy S) v CPC (M) ang nghin cu cng ngh sn xut b ca cao t du c, cht to v xaffron, v cay chilli, cht to mi cc loi, Mc khc, CNSH cng nh khoa hc ht nhn, bn cnh nhng ng dng cc k to ln cho li ch v pht trin ca loi ngi, c th mang li nhiu him ha khng th lng trc hu qu nh nhng lo lng v sn phm cng ngh gene, vn nhn bn, v kh sinh hc, mi trng, a dng sinh hc, Chnh nhng iu , quan h quc t v khu vc khng ch l s chuyn giao cng ngh pht trin khoa hc v cng ngh ca mi quc gia m cn to cho s pht trin bn vng ca th gii.
2. Quan h quc t v khu vc trong cng ngh sinh hc

2.1. Hng tip cn hp tc Hp tc quc t v khu vc l mt trong nhng bin php thc y chuyn giao CNSH v bin nhng kh nng ca n thnh hin thc cc nc ang pht trin. S hp tc quc t v khu vc c thit lp nhm nghin cu nhng vn chung, tin hnh nhng d n lin doanh nghin cu v em li nhng kt qu c th p dng c cho cc nc trong khu vc hoc tiu khu vc. Vic hp tc cng c th li cun c c cc quc gia cng nghip pht trin trong khun kh cc tha thun tay i, cng nh cc t chc t nhn, k c cc cng ti a quc gia. Cc lin doanh v cung cp kinh ph ca cc t chc t nhn hay nh nc v cc t chc quc t cho
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cc d n cng s gp phn chia s cng ngh v gia tng trao i kinh t gia cc nc pht trin v ang pht trin. Thi gian va qua, cc nc ASEAN thnh lp tiu ban CNSH (SCB) thuc y ban Khoa hc v Cng ngh (COST) ca ASEAN. Trng tiu ban SCB nhim k ca phin hp ln th 20 l GS Abdul Latifibrahim, MOSTE Malaysia. Cho n ngy 15/6/1999 do PGS.TS. Nguyn Vn Uyn, vin Sinh hc Nhit i thnh ph H Ch Minh m nhn. Hin nay c giao cho PGS. TS. L Trn Bnh vin Cng ngh Sinh hc H Ni m nhn. SCB c 5 ni dung chnh u tin nghin cu: - Pht trin thuc cha bnh, kt chn on v vaccine - Ci tin v sn xut vt liu sinh hc cho nng nghip v cng nghip. - ng dng CNSH nhm ci thin cht lng v sn lng cy trng vt nui v cc sn phm ca chng. - Thit lp qui m pilotte v iu khin bng vi tnh cc bnh phn ng sinh hc. - Bo v a dng sinh hc v mi trng. Cc d n c trin khai gia SCB vi cc nc trong khu vc. V d: + SCB vi Singapore: v sinh vt chuyn gene GMOs + SCB vi n : . Cng ngh sinh hc thc vt, nui cy m nhm ci bin ging cy trng v s dng c hiu qu ti nguyn sinh hc. . Cy chuyn phi ng vt tp trung trn i tng l ging b tht v b sa. . Cng ngh thng tin sinh hc. + SCB vi Hn Quc: Trin khai v bn cng ngh sinh hc ca cc nc ASEAN c s tham gia h tr ti chnh ca ESCAP, nghin cu v a dng sinh hc. Brazil cng ti Vallec Nordeste SA (Montes Claris, Minas Gerais) k hp ng vi vin Merieux ca Php v vi So Paulo Based Instituto Veterinario Rhodia-Merieux (Paulina), nhm s dng CNSH vo vic ch vaccine, thuc th y v huyt thanh ngi. Nhng sn phm ny s c bn trong nc. Tng s vn u t tr gi 70 triu USD. Vallec Nordeste SA ang sn sng hon thnh mt d nh c SUDENE (T chc Pht trin ca ng Nam Brazil) tn thnh vi mc ch to vaccine chng bnh un vn, bch cu, ho g v huyt thanh chng nc c rn.
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c bit s hp tc gia Brazil v cc nc ang pht trin v d n sn xut nng lng kt hp vi thc n. Chng hn: Brazil vi Senegal: Sn xut methan t b chui, methan ha v ci tin vic sn xut nng lng v thc phm. Brazil vi chu Phi: hp tc v m rng vic sn xut v s dng ci, ch to than, lm kh t t b hoa mu, xy dng nh my chng ct nh sn xut ethanol t r ng ma v cao lng ngt hoc t dch thy phn ca tinh bt Sn, sn xut methan t b hoa mu, phn gia sc v cht thi gia nh, phn gii bng enzyme v ch to sinh khi (biomas) t nguyn liu ligno-cellulose. Brazil vi n : Sn xut v s dng nng lng nh sn xut ethanol v biogas, s dng cht thi nng nghip, lm nghip v bo lc bnh, ti to rng vi nhng cy chng ln v pht trin cc h thng kt hp nng nghip vi nng lng (La Rovere, 1985) Cng ti pht trin quc t Hoa K (USAID) v qu Rockefeller bo lnh cho T chc Bt v li gi l cng ti Pht trin Ti nguyn (RDF) k t nm 1984, RDF tham d vo mt s cng ti thuc ngnh CNSH trong nhm CNSH quc t (IBG). Mc ch hp tc gia cc cng ti RDF v IBG l thu ht nhng hi vin a phng thuc cc nc ang pht trin vo cc d n chuyn giao cng ngh. Nhng d n ny cng c mc ch thng mi v th trng quc gia ang pht trin rt quyn r i vi cc cng CNSH. Chng hn, cc chuyn gia khoa Thng mi Hoa K tuyn b rng: cc nc Trung ng v Bc Phi l th trng c li nht cho cc sn phm CNSH. H d kin rng, cc quc gia xut khu du trong nhng vng ny b vn khong 50 t USD cho s pht trin nng nghip ca h trong nhng nm sp ti vi s tin cy sau ny ca ngnh CNSH (Zolty, 1987). V vy ngi ta hi vng rng, mc tiu tng qut ca s u t l vic chuyn giao k thut v kin thc nhm to iu kin cho cc nh khoa hc v k thut ca cc nc ang pht trin sng to v ci thin nn cng nghip ring ca h. Vic ch to vaccine chng l mm long mng Boswana vi s cng tc ca vin Merieux Php, chng trnh tim chng chng vim gan B (hepatit B) Senegal v i Loan, cng cn mc ch ngn cn bnh ung th gan s khi, vi s gip ca cc nc pht trin v cc c quan c thm quyn ca cc nc ny; vic to dng cy c da v xy dng cc n in mi cho loi cy ny Cte dIvoice, Malaysia v Indonesia lin doanh vi cc t chc nghin cu v pht trin ca Php (ORSTOM v IRNO) hoc cc cng ti a quc gia.
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S hp tc nghin cu v pht trin gia cc nc cng nghip ha v cc nc vng ng Nam ha hn kh nng tin n nhng k thut tin tin nht. Chng hn, khi hip c cu tr hai bn v nghin cu v pht trin trong ngnh cng ngh cao cp c k gia b Ngoi giao Hoa K v Thi Lan nm 1985, ngnh CNSH c nu ln nh mt lnh vc quan trng nht, vi tin u t l 50 triu USD (Yenchinski, 1987). Singapore ngnh CNSH v chn on bnh, lin doanh u t gia cc phng th nghim nghin cu Biotech (Marylan, USA) v cc nh u t a phng nhn thng tin t cc cng ti m v min dch hc, ha min dch Hybrodome v k thut di truyn. N s sn xut v bn nhng mu th nghim chn on virus HTLV v HIV (AIDS). Biotech Pte Ltd. ca Singapore mua bn quyn vaccine chng vim gan B theo k thut di truyn ca cng ti M Morck Co. Inc v bn cho cc quc gia chu , Hng Kng, Brunei, Myanmar, cng c k hoch pht trin nhng kt chn on bnh k c AIDS. c bit dng kt hp ca cc t chc nghin cu v gio dc i hc s c khuyn khch, vic thnh lp mng li quc t CNSH theo sng kin ca Php, vic hp tc lin quan n 3 lnh vc: s dng sinh khi thc vt, c nh N2 sinh hc v nng lng sinh hc. Cc nc ang pht trin chu u hp tc v CNSH vi tiu khoa hc v cng ngh cho s pht trin ch yu trong lnh vc nng nghip nhit i v y t. chu M Latinh, pht trin lin hp (UNDP) thnh lp chng trnh a phng ca ngnh CNSH, hoch nh v b sung nhng k hoch nghin cu v pht trin a phng, mc phng th nghim sn xut th v qui m cng nghip c lin quan n nhng vn a quc gia. UNESCO v UNIDO (T chc Pht trin K ngh Lin hip quc) gm cc quc gia Argentina, Chil, Colombia, Costa Rica. Cuba, Guatemala. Mexico, Pru, Uruguay v Venezuela c chng trnh Pht trin cng ngh v ng dng cng nghip CNSH. Nhng d n c thc hin nh chn on bnh trypanosoma v leishmania trong nhng a phng c bnh khc nhau ca chu M Latinh. S pht trin ca nhng th nghim chn on vi bnh virus thc vt, phn gii bng enzyme cc ch phm cng nghip thc phm, sn xut cng nghip penicilline-amidase v s dng chng tng hp 6-amino-penicillanic acid (6-APA), pht trin h thng chn on kim mi c nh du vi bnh st rtMi d n c Gim c iu hnh v 3 n 6 nc tham gia. Nhiu d n c lin quan n vic nhn ging trong ng nghim v k thut di truyn thc vt c chun b t nm 1988.
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Vo thng 4 nm 1987, trung tm CNSH Brazil-Argentina c thnh lp. Ln u tin chu M Latinh hai quc gia cng nhau thnh lp trung tm nhm mc ch pht trin v cng iu hnh nghin cu khoa hc v u t vo lnh vc CNSH. Cc nh cm quyn quc gia c hai nc nhn mnh rng ngnh CNSH khng nhng c s gip ca Brazil v Argentina m cn c nhng quc gia khc ca lc a cng gii quyt nhng kh khn do s ph thuc vo cc trung tm ln v p ng vi nhng thch thc gia tng trong lnh vc cnh tranh kinh t quc t gay gt ny. Trung tm hai quc gia ny t chc li, c s tham gia ca t chc nh nc, cc trng i hc, cc trung tm nghin cu v cc hi ng ca 2 nc tham gia vo vic tm kim cc gii php mi v pht trin nhng loi thuc mi, thc phm v nng lng. Hai bn tha thun thnh lp mt trng CNSH nhm o to cc chuyn gia trung v cao cp. Thng 8 nm 1987, trung tm Brazil-Argentina nghin cu v pht trin thuc cc lnh vc sau: ci thin v i mi cc vaccine chng un vn, bch hu, ho g; chn on v sn xut vaccine vim gan B, sn xut khng sinh bng enzyme; ci thin cy lng thc qua ng CNSH. Vic hp tc gia cc quc gia ang pht trin v cc quc gia pht trin hu nh c chng minh. C hai nhm quc gia u quan tm n hai sc thi ch yu ca CNSH v tc ng x hi v s tn ti tnh a dng di truyn. 2.2. Vai tr ca cc t chc chnh ph quc t Nhng thnh cng t c v thng mi v s vn u t khng l trong cc lnh vc khc nhau ca cng ngh sinh hc lm cho sc sng ca cuc cch mng CNSH c tng tin v khng th o ngc. Ch ring lnh vc nng nghip v cng nghip thc phm doanh s cc sn phm thu c do p dng CNSH mang li c th ln ti 50 n 100 t USD nm 2000. u th v li ch ca cuc cch mng CNSH ko theo cc hu qu x hi v c nh hng khng th ging nhau i vi tng nc v tng nhm t chc x hi trong cng mt nc.V vy, phi tm ra hng khc phc cc hu qu khng c li cho p dng CNSH v thit k chin lc thch hp phn tch mt cch cng bng cc ngun li mang li di s gip ca cc t chc chnh ph. Nhn thc v s cn thit bo v cc ging cy trng dn n vic k hip c ca Lin on quc t bo v ging mi (hip c UPOV International Union for the Protection of New Variteties of Plants) ca B, Php, Italia, H Lan, v Cng ha Lin bang c di s bo tr ca T chc S hu Tr thc th gii WIPO (World International Property Organization), mt c quan
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chuyn trch ca Lin hp quc v sng ch. y l kt qu ca mt s hi ngh quc t t nm 1957 n nm 1961. Hip c ny c sa li t nm 1978 v c coi l b khung lut thch hp bo v ging cy m cc thnh vin c th cng c thm thng qua php ch.. T nm 1978, quyn tham gia UPOV c m rng cho cc nc khng thuc chu u. n nm 1987, c 17 nc thnh vin l B, an Mch, CHLB c, Php, Nam Phi, Ty Ban Nha,Thy in, Thy S, Hungarie, Irland, Israel, Italia, Nht Bn, H Lan, New Zaeland, M v Anh. Mt khc, do s ph thuc ln nhau v khoa hc, cng ngh, kinh t v ti chnh ang ngy cng tng ln trn th gii, nn kh c nc no t c lp mnh. Mt khc s bt bnh ng v sc mua gia cc nc giu v nc ngho, gia cc nc ang pht trin v pht trin lm cho ngy cng kh t c cc tha thun cng bng c li cho c hai bn. Chnh v vy, vn bo v cc sng ch v CNSH l rt quan trng v c tip cn bng mt phng php thc dng. Hin c hai hng tip cn khc nhau v h thng quc t bo v sng ch cng ngh sinh hc. Ngoi hip nh ca UPOV ni trn c kin ca tiu ban chuyn gia v sng ch CNSH v s hu cng nghip ca WIPO: bo v cc qui nh hoc sn phm bng cch cp bng sng ch. Cc cy trng, gia sc, vi sinh vt c bin np u c bo v thng qua bng sng ch. Vn bo v ti nguyn di truyn thc vt v tip cn n tr thnh vn chnh tr ton cu. Kha hp u tin ca y ban lin chnh ph v ti nguyn di truyn thc vt hp vo thng 3 nm 1985 ti tr s FAO Rhoma, vic tham gia kha hp gm 67 nc thnh vin ca FAO v 27 nc thnh vin khc vi t cch quan st vin (a s trong l cc nc cng nghip) ko theo 74 nc thnh vin ng h cam kt, trong 57 nc khng bo lu v 17 nc c bo lu. Thng 3 nm 1985, mng li cng tc ging c thnh lp. Lin minh quc t v s pht trin (ICDA) chin dch ht ging. Ngoi ra, cc t chc quc t ng vai tr quan trng trong dch v t vn cho chnh ph nhm hnh thnh cc chnh sch v cc chng trnh quc gia trong ngnh CNSH nhm pht trin v sau , vic phi hp cc d n nghin cu hoc u t gia quc gia ang pht trin v cc quc gia cng nghip pht trin. Vic y mnh s tham gia ca cc nh nghin cu v cc nh k thut ca cc quc gia trong vic u t ny cng c nng lc ca cc quc gia trong vic nghin cu, hun luyn. V vy, t nhiu nm qua, cc chng trnh UNESCO, FAO v WHO pht trin v m rng hp tc quc t v vi sinh vt ng dng v CNSH trong lnh vc y t nng nghip v chn nui.
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Chng hn, nm 1962, UNESCO ti tr cho s thnh lp t chc nghin cu t bo quc t (ICRO). Nm 1972, tip theo hi ngh Lin hip quc t v con ngi, mi trng (Stockholm) thng 6 nm 1972, UNESCO phi hp vi ICRO v UNEP, chng trnh mi trng gi gn bo v ti sn di truyn gm cc ngun vi sinh vt v lm cho cc nc ang pht trin c th tip cn c nhng cng vic . Giai on u tin trong vic thnh lp mng li MIRCEN l s thnh lp trung tm T liu th gii v cc vi sinh vt Brisbane, Australia. Gn y, trung tm ny chuyn sang Nht Bn, c s MIRCEN khc t ti Bangkok (vin Nghin cu Khoa hc v K thut Thi Lan) cho vng ng Nam , ti Osaka (vin CNSH v i hc Osaka) v Saitama, Nht Bn (Rikagaku phng Sinh hc), ban Thng tin, Bambey, Senegal (trung tm Nghin cu Nng nghip quc gia) v Nairobi, Kenya (b mn Thc vt Th nhng , i hc Nairobi) cho chu Phi, Porto Alegre, Brazil (Instituto de Pesquisas Agronomicas) Tucuman, Argentina (Plata Piloto de Prosesos Industriales Microbiologicos, PROIMI) cho Nam M, Cindad Guatemala cho Trung M v Cairo, Ai Cp (i hc Ain Shams cho cc nc A Rp). MIRCEN Hawaii (d n NifTAL, khoa Nng nghip Nhit i, i hc Hawaii) dnh hu ht hot ng ca mnh cho vic c nh N2 ca cc loi rau nhit i. Mng li MIRCEN cn c cc trung tm i hc Maryland, M (b mn Vi sinh), cc i hc Waterloo v Guelph, Ontario, Canada, cc i hc Kent v Centerbury, nc Anh, vin Karoliska (Stokholm, Sweden) h tr v nng , k c MIRCEN Php (Centre de transfort en Microbiolgie, Touluse) ni c nhiu vin v phng th nghim tham gia. Nm 1981, c quan Pht trin K ngh Lin hip quc (UNION) thnh lp trung tm quc t K thut di truyn v CNSH (ICGEB) Irieste v New Delhi vi ngn sch 40,7 triu USD. Ngun qu qu hn hp, song cc t chc chnh ph quc t cng ng gp ng k h tr cho cc quc gia ang pht trin trong lnh vc CNSH. 2.3. Vit Nam vi hp tc quc t v khu vc trong cng ngh sinh hc Vit Nam l mt nc nng nghip ang trn bc ng cng nghip ha v hin i ha, cng nm trong bi cnh chung ca ton khu vc. Cuc cch mng cng ngh sinh hc s l ng lc gp phn to ln i vi s pht trin kinh t-x hi. Thy c tm quan trng , Chnh ph Vit Nam ra ngh quyt 18/CP ngy 11 thng 3 nm 1994 v phng hng pht trin khoa hc v cng ngh nc ta. Trong , Nh nc nhn mnh vic hp tc quc t v khu vc trong khoa hc v
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cng ngh, c bit l trong CNSH. Vit Nam l thnh vin chnh thc tham gia vo Hip hi cc quc gia ng Nam v Khoa hc v Cng ngh vo nm 1995. Tun l Khoa hc v Cng ngh ASEAN ln th V c t chc ti th H Ni t ngy 5 n ngy 15 thng 10 nm 1998 vi ch Khoa hc v Cng ngh - ngun ng lc hng ti pht trin bn vng ca ASEAN. Vi s tham gia ca hng trm i biu v trn 500 nh khoa hc Vit Nam, cc nc thnh vin ASEAN khc cng nh cc nc i thoi ca ASEAN mang m du n Vit Nam vi tinh thn xy dng ASEAN thnh cng ng cc quc gia pht trin bn vng, hp tc v ng u, c cng ng cc nh khoa hc trong nc v quc t nh gi cao. Mt s kin c coi l hot ng v khoa hc v cng ngh c nhiu ngha v ln nht v qui m t trc n nay. Vit Nam m rng hp tc vi Cuba trong lnh vc qun l khoa hc v cng ngh, mi trng, cng ngh thng tin, cng ngh sinh hc v o to cn b (thng 9 nm 1998). Chng ta cng hp tc vi Lin bang Nga trong vic nghin cu khoa hc th nghim nhit i, nghin cu hu qu v sinh thi v y sinh hc ca chin tranh ha hc do Hoa K tin hnh Vit Nam. Vi Hn Quc, chng ta hp tc v k thut thnh lp trung tm hp tc cng ngh Vit Nam-Hn Quc (ViKotech) vi tng s vn l 2.880.000 USD, vin tr khng hon li ca Chnh ph Hn Quc, Vit Nam ng 500.000 USD. Hp tc vi Hoa K: trao i v k thut v hp tc trong lnh vc khoa hc vt liu, cng ngh thng tin v cng ngh sinh hc vo thng 1 nm 1998. Hp tc vi Thy in: c chuyn gia Thy in vo Vit Nam o to cn b v cng ngh sinh hc, cung cp mt s trang thit b c bn nhm tng cng c s vt cht cho v sinh dch t hc H Ni, xy dng phng th nghim chun thc quc gia v vi khun ng rut, vaccine l Shigella, c ch gim nh tnh hnh khng thuc Vit Nam, ci thin cy trng rng, k thut nui cy m v t bo, k thut chun on nhanh k sinh trng st rt. Vit Nam ngy cng ch trng n quan h hp tc quc t v khu vc chuyn giao cng ngh, o to i ng nghin cu khoa hc trong lnh vc cng ngh sinh hc, thu ht vn u t pht trin khoa hc v cng ngh gp phn vo vic pht trin kinh t x hi ca t nc. S hp tc cng ngh sinh hc trong khu vc v quc t khng nhng l bin php thc y chuyn giao cng ngh pht trin khoa hc v cng ngh trong mi quc gia m cn nhm nghin cu nhng vn
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chung; tin hnh nhng d n lin doanh nghin cu v em li hiu qu c th p dng chung cho cc nc trong khu vc. Ngoi ra, hp tc quc t v khu vc khng lm trm trng thm s chnh lch gia cc nc trong lnh vc khoa hc v cng ngh, ngn nga v gii quyt nhng hu qu nguy hi cho s pht trin ca cng ngh sinh hc nh: cc sn phm t vic chuyn gene, vn nhn bn, mi trng, a dng sinh hc, v kh sinh hc, Vn kin Hi ngh ln th 7 BCHTW ng kha VII (thng 7 nm 1994) ca ng ta cng nhn mnh tm quan trng ca CNSH trong vic pht trin nng, lm, ng nghip, cng ngh ch bin thc phm, dc phm v bo v mi trng sinh thi. Cc ch trng c th l: Thc hin c cu cng ngh kt hp nhiu trnh , cc gii php v cng ngh phi ly hiu qu kinh t-x hi gn vi bo v mi trng sinh thi lm tiu chun cao nht. Hng chnh i mi nhanh cng ngh v nhp cng ngh tin tin v hin i, ng thi khuyn khch, ci tin v sng to cng ngh mi. Trong cc d n c vn u t nc ngoi, cn ch yu t chuyn giao cng ngh. Ch trng cc cng ngh i hi sut u t thp, thu hi vn nhanh, c kh nng to thm nhiu ch lm vic trc tip v gin tip, tranh th i mi cc thit b, hin i ha cng ngh trc ht mt s khu c ngha quyt nh i vi vic nng cao cht lng sn phm, nht l hng xut khu mt s ngnh c tc dng trc tip vi vic nng cao trnh cng ngh ca nhiu ngnh khc v mt s lnh vc v a bn i hi sm vn ln ngang trnh vi khu vc v quc t,
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4. 5.
6.
TI LIU THAM KHO L Trn Bnh, 1999. Hp tc v cng ngh sinh hc ca cc nc ASEA. Bo co khoa hc Hi ngh Cng ngh sinh hc ton quc ngy 9-10 thng 12 nm 1999. Nxb Khoa hc v K thut H Ni, tr: 52-56. Trng Vn Lung, 1995. Chuyn cng ngh sinh hc. T sch i hc Khoa hc Hu Nguyn Vn Uyn, Nguyn Tin Thng, 1996. Nhng kin thc c bn v cng ngh sinh hc, Nxb Gio dc H Ni. Tp ch Hot ng khoa hc,S 2, S 21/1999. Vn kin Hi ngh ln th 7 BCHTW ng, kha VII, trang 53-64. Albert Sasson,1988. Biotechnologies and development Cng ngh sinh hc v pht trin. Ngi dch: Nguyn Hu Thc, Nguyn Ln Dng v mt s dch gi khc. Nxb Khoa hc & K thut H Ni.
CNSH vi vic pht trin bn vng
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Chng XII:
Nhng nh hng chnh v s pht trin CNSH hin nay th gii cng nh Vit Nam.
Trong lnh vc cng ngh sinh hc, vic nghin cu khoa hc cn c nhng nh hng khng ngng khm ph nhng iu mi l xy ra trong c th sinh vt m con ngi chng ta cn bit nhm khng ngng phc v cho i sng kinh t x hi ca con ngi hin nay. Sau y l mt s nh hng chnh v s pht trin cng ngh sinh hc hin nay trn th gii cng nh Vit Nam.
1. Gii m b gene ca cc sinh vt khc nhau v ngnh genome hc

Vi s tin b ca my mc v thit b k thut, t ch c trnh t nucleotide ca mt on DNA c tin hnh theo phng php th cng mi tun mi ngi ch thc hin c mt vi phn ng vi nng sut 300 bp/phn ng, n nay vi h thng my mao mch c th xc nh t ng ng thi 96 phn ng vi di trn 1000 bp/phn ng th cc n xc nh ton b trnh t nucleotide ca b gene nhiu sinh vt c thc hin, trong c n xc nh trnh t genome ngi di 3,3 t nucleotide hon thnh vo thng 2 nm 2002, ng 50 nm sau khi Watson v Crick pht minh ra m hnh cu trc xon kp ca phn t DNA to ra bc thay i cch mng trong nghin cu sinh hc phn t. n nay c ti hng chc n xc nh trnh t nucleotide b gene ca nhiu sinh vt c hon thnh. Thnh tu v gii m b gene ngi (99% genome ngi c c vi chnh xc 99,99% vi 30.000 gene) v nhiu sinh vt khc nh cy la nc (Oryza sativa L. l 50.000 gene), Escherichia coli, Saccharomyces cerevisiae, C. elegan, Drosophila melanogaster v bn nhp ton b h gene ca vi loi khc nh C. briggsae, D. pseudoobscura, chut. Vit Nam cng gii m thnh cng virus gy dch cm gia cm H1N5 Genome hc i vi cc nc ang pht trin nh cc nc trong khu vc ASEAN trong c Vit Nam nh hng vo vic xc nh cc c im ca h gene cc nhm dn tc ang sinh sng trn cng lnh th v cng khu vc, phc v cho vic khm cha bnh v ng dng vo lnh vc x hi hc.
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2. Phn tch tng th cc bin ng ca h protein t bo v ngnh protein hc

Mt ngi trong cuc i phi tri qua mt qu trnh pht trin t khi cn tr n lc v gi, lc m au, khi khe mnh. Trong qu trnh b gene lun lun c nh, cn protein th li thay i ty theo tng trng thi, giai on pht trin ca con ngi. Nm c protein c ngha l nm c c ch iu khin ca mt chu trnh sng trong con ngi chng ta. Bn gene ngi mi ch l bc i u tin trong qu trnh nghin cu. Bc tip theo, cc nh khoa hc theo ui mt cng trnh c th ko di trong nhiu thp nin mang mt ci tn cn rt l lm ngay trong gii khoa hc: Proteomics- nghin cu b gene ca con ngi. Kh khn nht ca cc khoa hc gp phi trong khi nghin cu protein l qu trnh bin i phc tp trong t bo, ch mt gene thi c th tng ng vi 20 protein khc nhau. GS. Angelika Goerg (trng i hc Tng hp Munich-c), mt trong nhng nh khoa hc i tin phong trong cng cuc nghin cu protein cho bit, trong khi b gene khng thay i th thnh phn ca cc protein li bin i theo tui tc, tc ng ca mi trng ngoi cnh, thuc men v bnh tt. Mi loi t bo trong tng s 270 loi t bo khc nhau ca con ngi khi dch b gene (gm t 30.000 n 35.000 gene) lun lun ging nhau u to nn nhng protein hon ton khc nhau. Chng ta c th ly nhng thc t sau y lm v d cho tnh bin i ny: mt con nhng v mt con bm; mt bn tay con tr v mt bn tay ngi gi hoc mt ht ging v mt bng hoa. Tt c nhng cp u c cng mt b gene, nhng li mang nhng protein, hoc ni chnh xc hn: nhng nhm protein khc nhau. Cho n nay, con ngi cn bit qu t v hn 400.000 protein ca chng ta S lng protein nhiu nh vy c hnh thnh khi m di truyn c dch ra theo nhng trt t sp xp ca cc acid amin trong protein. Tuy nhin, trt t sp xp cc acid amin c m t trong khng gian mt chiu hon ton khng ni ln iu g v chc nng ca protein c. Yu t quyt nh l s sp xp ca chng trong khng gian ba chiu.Cc hnh thc sp xp khc nhau ca protein, hnh xon, hnh cun li thnh b hoc hnh gp np (cu trc bc I, bc II, bc III, bc IV) quyt nh chc nng ca n. Mt cu to theo hnh gp nhng b sai lch i ca protein c th gy ra nhng hu qu gh gm, v d nh bnh b in. Mi n nm 1995, nh khoa hc ngi Anh Frederick Sanger ln u tin mi tm ra cu to ca protein trong insulin. GS. Sam Hanasch, mt chuyn gia v ung th ca trng i hc Tng hp Michigan (M) v ng thi l ch tch T chc Quc t Nghin cu protein ca ngi (HUPO) c thnh lp nm 2001 cho rng, c l
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trong sut cuc i mnh, con ngi phi c n hng triu protein khc nhau. ng nhn xt: Nhiu phng th nghim nghin cu protein t nhiu nm nay, nhng mi gn y mi nhn thy rng, gi y vi s pht trin ca cng ngh th thi i ca protein mi bt u. Hin nay, cc trung tm nghin cu ca cc trng i hc v cc tp on kinh t ln, mi ngy, hng ngn protein c em ra m x phn tch. Nhng thng s ca chng c so snh vi thng s ca ngn hng gene. Chng ta cng bit, trnh t nucleotide ca b gene mi l s khi u trong qu trnh nghin cu v b gene. Vic nghin cu ton b h protein do cc gene m ha v iu khin sinh tng hp trong tng giai on pht trin v trong tng trng thi sinh l, bnh l ca sinh vt v c bit l ca con ngi ang l lnh vc thu ht s u t qui m ti hng trm t USD, nht l trong nghin cu h protein ca ngi. Vi cc k thut sc k, in di trc y, ngi ta ch nghin cu c tng loi protein ring r. Hin nay, khi phi hp sc k a chiu v khi ph, ngi ta c th phn tch c 5.000 loi protein cng mt ln v kt qu cho php chn on sm nhng bnh him ngho nh ung th mu, Mc tiu lu di ca nhng nghin cu proteomics l gii m c chc nng sinh hc ca h gene. Song, trc mt nhng bin i hot ng ca cc nhm gene trong iu kin bnh l s cung cp thng tin cho vic chn on sm, phng tr v iu tr nhiu loi bnh. Bi v, 98% cc loi bnh tt l do protein iu khin. Gene v s sai lch v gene ch gy ra khong 2% tng s cc loi bnh tt m thi. Cho d l viagra hay aspirine hin c n hn 90% cc loi thuc tc dng n protein. Hin nay, mt trong mc tiu ng dng hng u c t ra l da trn nhng hiu bit mi nht v protein tm kim nhng loi bit dc mi v c cc cng ty Dc cht ch trng bo v b mt thng qua ng k pht minh sng ch. GS. Patterson Gim c cng ty Celera ni ting trong lnh vc gene cho bit, cc nh khoa hc ca ng ang mt mt mun chn on c rt sm bnh ung th hoc co tht c tim da trn vic phn tch protein trong nhm mu, tm kim nhng bit dc hu hiu c kh nng n bt cc protein gy bnh, mt mt hp tc vi cc chuyn gia my tnh ca tp on Compaq v chic my tnh Red Storm khng l ca h ang ngy m tnh ton tm hiu nhng b mt trong th gii protein.
3. Cy trng chuyn gene

Nh trn phn ba, chng VIII, mc 2.4. Ch bin thc phm chuyn gene chng ti c dp cp n vn ny. y, v l s nh hng chung ca th gii nn chng ti li nhc n ln na.
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Gia nhng nm 1990, cng ngh gene bt u a vo hn 50 sn phm mi c ng dng 13 nc vi din tch 52,6 triu ha nm 2001, tng 50 ln so vi nm 1996. Trong nhiu nht l u tng 34,9 triu ha, bng 64% din tch u tng th gii, ng 6,1 triu ha v 3,3 triu ha cy ci du. Ngi ta ra c hn 10 ging cy trng mang gene mi. n nm 2003, sau cha y 15 nm, s din tch trng cy chuyn gene ln n 67,7 triu ha.Trong nm 2004, din tch cy trng chuyn gene tng 20% so vi 15% nm 2003 v t 81 triu ha. D tnh vo nm 2010, din tch cy trng cng ngh sinh hc trn th gii s tng ln n 150 triu ha vi kong 15 triu ngi trng ti 30 nc trn th gii (nm 2004 c 8,25 triu nng dn ti 17 nc trng cy chuyn gene). Nhng gene g c a vo cy trng? l nhng loi gene tng cng kh nng khng su bnh nh gene khng su nhm cry/VIP, gene khng virus nhm CP/Nbi, gene khng thuc dit c nhm bar. n nay ngi ta ang tm cch a gene sn xut vaccine, gene sn xut dc cht vo cy trng t cy lng thc thc phm thnh cy sn xut dc liu c gi tr kinh t cao hn. Trong s hn 50 loi cy trng mang gene chuyn ang c th nghim th cy bng vi khng su, cy u tng, cy ng khng su, khng cht dit c chim tng s trn 90% din tch gieo trng ni trn. nc ta, nhng nghin cu to bng khng su, chu hn, to la go giu -caroten ang c tp trung gii quyt. Cng vi tin b trong nghin cu, cng tc chun b vn bn php l cho vic nghin cu v s dng cc sinh vt chuyn gene cng ang c chun b rt thn trng, ph hp vi tnh hnh trong v ngoi nc.
4. Sn xut v ng dng chp DNA

Affymetrix l cng ty hng u th gii trong vic sn xut cc loi chp DNA l mt mnh mng lin kt c kch thc 2040 mm c in trn bng cc on DNA dng nhng im chm vung cc nh, v d chp genome ngi c in 20.000-25.000 gene V c hnh dng ging nh mt con chp vi nhng chm DNA thay cho chm in t gn trn mt lt thy tinh cc nh nn c gi l chp DNA. Ta bit rng, trt t sp xp base ca mt si DNA s tit l cu trc si DNA kt hp vi n. Nh khoa hc tr Stephen Fodor tin rng c th gii m DNA bng cch cho n lin kt vi DNA bit trc cu trc gn trn chp, nh t bin v trnh t sp xp cc base s c pht hin v bit bnh g hay cch iu tr. Mt con chp c th c hng ngn gene cng lc. Nm 1993, Fodor v 9 ngi khc ca Affymax thnh lp cng ty Affymetrix (California, M) thc hin tng ny. Khng ch Affymetrix cc con chp mi ra i m i hc Stanford, phng th nghim Palo Alto v Sunnyvaie (bang California)
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cng nghin cu ch to loi chp ny. Mt robot en sc vng ang chm hng ngn m DNA ln mt lt ct thy tinh cc nh, cc nh nghin cu nh mu xanh d quang cho phn t RNA (RNA gi nhim v truyn thng tin di truyn ca DNA n ni sn xut protein) ca t bo ung th, cn RNA ca t bo thng c nh mu d quang. Khi c tri ln con chp, cc phn t RNA ny bm vo cc on gene tng thch vi chng; gene hot ng mnh hn trong t bo ung th s chp xanh, gene hot ng trong t bo lnh ln chp .Chnh nhng bin i hot ng ca gene trong t bo ung th s l mc tiu cho nhng loi dc phm mi tr ung th. Hot ng ca gene cng cung cp v trng thi ca bnh ung th ang di cn nhanh, cn iu tr ngay hay bnh ang thoi trin ch cn theo di k. Nh vy, khi lai mnh mng chp ny vi sn phm phin m ca genome c th cn nghin cu cc chm DNA s i mu tng ng vi mc hot ng ca nhng gene trong c th. trng thi v thi im nghin cu cho php kt lun v tnh trng bnh l ca i tng nghin cu. Bc s c th dng mt con chp DNA chn on xem gene ca bnh nhn c mang mm mng ca bnh tim hay bnh alzhemer khng. Hoc gi bnh nhn mc bnh ung th th con chp s cho bit mc nghim trng ca bnh v xut loi thuc hiu qu nht. Bnh nhn c th ri phng mch bc s vi danh sch bnh c th xy n cho mnh trong vi nm ti, km theo l cc thay i v li sng, ch n ung v mt s toa thuc nga bnh. Lc con ngi c th kim sot phn no nh mnh ca mnh hn sn trong gene. Kinh ngc hn na l con chp c th d on v s kho lo hay tr thng minh ca mt hi nhi mi cho i. Ngy nay s nghin cu cc con chp sinh hc ang c tin hnh ro rit bin nhng ng dng trn thnh s tht.Cc k s chp sinh hc ti Affymetrix ang thi ua vi cc ng nghip bn ngnh bn dn nhi nht n 400.000 chui DNA khc nhau ln mt con chp c th gii m cho on DNA di 100.000 n v. Vi tin b , ngi ta hi vng mt th h chp mi s gii m di truyn ca mt ngi ch sau mt m. Ngoi vic c gene ngi, con chp ca cng ty Nanogene San Diego (Mi) cn tm kim du hiu nhim khun trong mu trong vng 15 pht. K thut ny cng dng pht hin mi loi vi khun c y hc bit n v c th ng dng rng ri cho m, dch nhy cng nh trong ngnh tm cht nhim bn nc v thc phm. Chp DNA cng ang dn dn tr thnh cng c chn on trong cng nghip ln men vi sinh vt, trong y hc d phng, trong kim dch ng thc vt v v sinh an ton thc phm, trong theo di mc
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nhim mi trng, vi tnh nng nhanh nhy v t ng ha cao. Hin ti th gi thnh ca mt con chp cn cao, nhng khi nhu cu s dng cng pht trin th gi c s gim dn.
5. Sinh tin hc (bioinformatics)

Cng ngh thng tin l ngnh khoa hc nghin cu vic ng dng my tnh in t v k thut thng k vo vic qun l v x l cc thng tin sinh hc. Trong n genome cng ngh sinh tin hc (CNSTH) bao gm c vic xy dng pht trin cc phng php tm kim khai thc nhanh ngn hng d liu, phn tch trnh t v cu trc ca DNA v protein. STH cng bao gm vic thu thp s liu, phn tch, qun l tp s liu v tm kim khi cn. STH ang ci tin phng php x l phn tch s liu, ci thin kh nng d on vng hot ng, vng ngng ngh ca b gene, ci tin kh nng phng on phn ng t bo i vi tc nhn ngoi sinh, thit lp nn cc cu trc phn t protein c hot lc cao v nh hng phn ha t bo mt cch hiu qu Hin ti ba ngn hng d liu gene ln nht th gii l World Gene Bank (M), EMBL (Chu u) v JDDB (Nht Bn) lu tr trn 9 t d liu v gene. Ngoi ra cn c ngn hng d liu v protein ca Thy S (Swiss Protein Database) cn lu gi ngoi thng tin v trnh t acid amin, cc tnh cht v chc nng sinh hc cn c c phn m hnh cu trc phn t ca cc loi protein na. Cc ngn hng trn u thuc loi cng cng, mi ngi u c th s dng min ph. Ngnh ngh trong STH bao gm chuyn gia sinh tin hc, chuyn gia lp trnh STH v khai thc ngn hng d liu, cui cng l chuyn gia qun l CNTTSTH. Mt s thnh tu ni bt ca STH c th l: chng trnh NMR a chiu thit lp cu trc khng gian protein; chng trnh FASTA so snh trnh t gene v protein ra i trc nm 1990 cho php so snh t ng, min ph trnh t mt on gene di khong 1.000 bp vi 9 t trnh t cng b trong vng vi chc pht; chng trnh BLAST, trung tm NCGR, chp DNA thit lp trc nm 2000 v gn y l n IBM Blue Gene c bt u, h thng chng trnh trn gi EMBOSS c bn, chp DNA b gene ngi c a ra th trng. n thi im ny c trn 60 cng ty ln chuyn dch v v kinh doanh trn lnh vc STH ang hot ng, trong phi k n Celera Discovery System tham gia vo n xc nh trnh t genome ngi. DNAStar hoc GCG l hai trong nhng cng ty thnh cng nht trong lnh vc cung ng phn mm phn tch gene; InforMax hot ng trong tp on Invitrogene.
6. Nhn bn ng vt
Nhn bn v tnh l k thut nhn s dng mt c th, mt mnh m hay mt t bo duy nht to ra hng lot c th hon ton ng nht
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v di truyn m khng thng qua qu trnh th tinh hu tnh. C s khoa hc ca vic nhn dng l tnh ton nng ca t bo (totipotency). Khi nim nhn dng thc vt tr thnh thng dng trong ngh trng trt cc loi cy n qu, cc loi hoa, cy cnh, cc loi cy cng nghip bng con ng nhn dng vi phu (micropropagation). Nhn dng ng vt mc d k thut nui cy m t bo ng vt ra i trc c nui cy m thc vt, nhng kt qu li ch mi hn ch vic duy tr cc dng t bo c kh nng phn chia lin tc v sn xut ra nhng loi thuc cha bnh qu him. Mi n nm 1997, Jan Winmut bng cch cy nhn t bo tuyn v vo trng cu th tinh b loi b nhn thu c phi v cy phi c con cu Dolly. Vic i nhn ca t bo trng cu bng nhn ca t bo tuyn v v vic nui cho t bo thay nhn thnh mt cm t bo phi bnh thng l mt bc tin rt xa ca khoa hc. Thnh tu ny mt ln na minh chng cho s ng n ca hc thuyt v tnh ton nng ca t bo. B my di truyn ca mi t bo ca c th u ging nhau. B quyt ca s thnh cng y l ngi ta chn ra iu kin thch hp cho cc nhn t bo bnh thng phn chia v lm nhim v ca nhn t bo trng, tc l to ra mt khi t bo phi. iu kin chnh l phn cht nguyn sinh cn li ca t bo trng khi b mt nhn. Thc ra, PGS.TS. Nguyn Mng Hng (cn b ging dy trng i hc Khoa hc T nhin thuc i hc Quc gia H Ni) t phng th nghim ca trng i hc Tng hp Lomonosov Moskva cng to dng v tnh thnh cng trn i tng con c chch, cch 20 nm trc khi con cu Dolly ra i.. Thnh cng trong vic nhn dng mt loi ng vt l s kt hp ti tnh ca 4 k thut: (1) nui cy t bo ring r tch t cc b phn khc nhau v d tuyn v, ty xng, biu b, (2) vi phu t bo loi b nhn ca t bo trng, thay vo l nhn ca t bo nui cy. (3) nui cy phi bt u t trng thi t bo trng ti phi a bo (4) cy chuyn phi vo d con ca ng vt mang, phi pht trin thnh thai v ra con vt non. Kt qu nhn dng ny thu c mt con vt ging m, nhng khng phi ging hon ton v DNA ca t bo cng c trong ti th cn li trong phn t bo cht ca trng sau khi loi b nhn. n nay tuy rng con cu Dolly qua i nhng c hng chc loi ng vt nhn bn v tnh nh b, ln, ch, mo, th, nga v c bit l chut chuyn gene,ch yu nh hng phc v y hc v cc nghin cu vai tr ca gene. Trn quan im kinh t th k thut nhn dng hng lot mt s ging cy trng hay vt nui siu sn u mang li hiu qu rt ln, nhng xt theo kha cnh an ton sinh hc th tim n mi him ha lm gim s a dng kiu gene s b n iu ha v thiu mt s phong ph a dng v tnh chng
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chu. Nhng ci li vn ang thng th. Bi l, nhiu ng vt hoang d ang c nguy c dit chng v nhng ng vt dit chng cn c cch phc hi li nh tng ca cc nh khoa hc Trung Quc ct ly mt mu da v ngm vo azote lng nhit -196 oC, mu vt lu gi c ton b h di truyn. Ngi ta cho rng, cch tt nht l ng lnh tinh dch con c v trng con ci tin hnh vic to thnh phi thai. Phi c ngm trong azote lng ch b m mang thai h (b m phi thuc loi tng cn). (xem thm trong chng XI: Bo tn v pht trin ngun gene qu him, mc 2, on: v ng vt trang 205).
7. Nui cy t bo gc (stem cells) Nhng nm gn y, cc nh khoa hc bt u khm ph nhng gene, protein v cch thc dn n kh nng ti sinh t nhin cc b phn ca ng vt. Alejandro Sanchez Alvarado i hc Salt Lake City (bang Utah-M) nhn xt: Con ngi c nhng gene m loi giun dp s dng ti to no, c bp, u b hy hoi. David Stoam H Purdue (bang Indiana-M) nghin cu Nng nc loi cc Xenopus laevis v cng nhn xt ging nh Sanchez Alvarado. Cc loi ng vt c kh nng c th d tr sut i mt t t bo gc mang ra s dng khi cn (giun dp, thy tc, cua,).Trong c th, t bo trng thnh c trong m t c kh nng bin ha thnh nhiu loi t bo khc nhau to nn hnh dng cc c quan m ch c t bo gc mi lm c vic ny. Nm 2000, Thomas Holstein (i hc Cng ngh Darmstadt, c) nhn dng mt loi thy tc tit ra protein Wnt vn l cc phn t ca ng vt c xng sng v nhn thy ch cn khong 10 t bo cng to thnh mt cc u mi.
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Peter Schultz (vin Nghin cu Scripps San Diego bang California, M) tm cch tng hp c mt s lng ln cht purine (gm nhiu loi phn t nh thng lin kt vi cc protein tc ng vo qu trnh pht trin ca t bo). H tm nhng purine no lm o ln qu trnh phn chia ng c ca chut v chn lc ra mt loi purine gi l hp cht myoseuerine c chc nng chia tch t bo t bo bt u phn chia. Trong khi , Mark Keating (i hc Y khoa Harwar bang Massachusette, M) li quan tm n mt gene gi tn l msx1 vn c chc nng sn xut mt protein kim sot hot ng ca cc gene khc trong c bp. Khi sa ging b ct mt u chn, chnh gene ny kch thch cho u chn mc li nh c. Gene msx1 cng gi vai tr to thnh cc chi ca phi thai chut. C l mt protein no ni b phn b ct ca sa ging c chc nng gi li cc mch tn hiu ng yn.
Nu nhn dng c protein ny, chc chn cc nh khoa hc c th to ra t bo gc bt k lc no v bt k ni no trong c th khi cn thit.
T b o g c -TtBO GC-lSTEM CELLS s em c e l s
Hnh XII. T bo gc
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Nh vy l, trong th k XXI, y hc s c c mt cng c cha bnh mi, hu hiu cho nhn loi, l liu php t bo. .Khi phi cn giai on rt sm mi c 8 t bo th mt t bo u c kh nng pht trin thnh mt phi hon chnh hoc phn ha thnh bt k loi t bo no ca c th sau ny. Nhng t bo ny, nh ta bit c gi l t bo gc nguyn pht. nhau thai mt s t bo cng cn duy tr c kh nng phn ha ton nng v c th nui cy thnh dng t bo gc th pht. T bo gc c dng thnh cng trong vic to ra gic mc mt ang dng trong vic iu tr ghp gic mc, c th nui thnh t bo c tim dng trong iu tr vt thng tim sau tai bin, c th thay t bo ty xng trong iu tr ung th muv hi vng trong tng lai khng xa c th gip phc hi c cc mch mu thn kinh b h hi trong iu tr bi lit hoc bnh alzheimer. Ngi ta hi vng c th nui t bo gc thnh cc loi c quan nh gan thnphc v vic cy ghp thay th c quan. Ngn hng t bo gc u tin ca nhn loi ra i Anh vi mc ch l thc y tht nhanh qu trnh nghin cu loi t bo thn k ny. Ngn hng ny ang l nim hi vng cho rt nhiu nh nghin cu v ngi bnh. T bo gc c kh nng cho php to ra mt c quan hon chnh. l t bo ES m ngn hng Anh ang c t nhng bo thai 6 ngy tui. Bng nhng k thut tin tin nht, ngn hng ny s trch c ES, sau nhn chng ln nh k thut ghp hay sinh sn v tnh.(Theo L Vn: Ngn hng t bo gc u tin trn th gii), Tp ch th gii mi, s 595, tr: 56-59, ra ngy 26/7/2004. Theo tin tc t Seoul, Hn Quc (12/2/2004), cc nh khoa hc Hn Quc nghin cu thnh cng vic nhn bn v tnh phi ngi. T 245 trng nhn thnh 30 phi. Phi pht trin t t bo gc v c kh nng phn chia thnh t bo c th. Theo bc s Moon, cc nh khoa hc Hn Quc khng c nh nhn bn v tnh hon chnh mt con ngi m trong qu trnh phn chia phi thnh cc c quan, cc nh khoa hc dng n trong vic cha tr mt s bnh him ngho. H ang th nghim thay th thn kinh ct sng cho mt ngi b h hng thn kinh ct sng. Trin vng ngi ny c th i li bnh thng m bao nhiu nm h nm nguyn ti ch.
8. iu tra hot cht sinh hc (bioprospecting)

iu tra thm d hot cht sinh hc trong sinh vt sng c th mang li li ch to ln v y dc hc v thng mi. ng nhin y l lnh vc u t mo him, c th tht bi v cng c th thu li nhun rt ln. V th, cc quc gia ang pht trin thng khng tim lc v ti chnh, v trang thit b v c bit v chuyn gia trin khai cng vic ny, mc d ti nguyn thin nhin v tnh a dng sinh hc ca cc nc
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ny ht sc phong ph v to ln. Doanh thu lnh vc ny qui m ton cu ln n 14 t USD hng nm. Hot ng iu tra thm d hot cht i hi s kt hp cht ch gia kinh nghim c truyn trong vic s dng cc ng vt, thc vt, nm v vi sinh vt lm thuc cng nh nhng trang thit b hin i ca phn tch ha hc v sinh hc. Cng vic thng bt u vi nhng nhm cy con c y hc c truyn s dng, sau m rng mt cch c h thng n cc khu vc vn Quc gia, tip n l cc h vi sinh vt sng trong cc h sinh thi c bit nh rng ngp mn, k sinh trn cy dc liu, trn rong, to bin, V phng php tin hnh trc tin l iu tra tm hiu kinh nghim s dng thuc ca ngi dn, sau tch chit, thng l cc loi dch chit, c a v phng th nghim tin hnh sng lc qua cc php th c trng cho cc nhm thuc nh khng khun, khng virus, cha st rt, cha ri lon thn kinh, cha bnh lao, c ch sinh trng ca t bo ung th. Rt c th phi phi hp nhiu phng th nghim t chc sng lc mt cch c h thng v c hiu qu. Nhng mu dng tnh s c phn tch ha hc nh sc k cao p, in di, khi ph cui cng i n hot cht. V d, gn y ngi ta tm thy cht chng HIV gi l conocurvone trong r cy Cotinus coggygria. Rt nhiu quc gia pht trin hoc cng nghip ha (NICs) v cc cng ti ha dc cht mun u t cho loi cng vic ny. V th, nhiu t chc quc t cho ra i nhng vn bn nu nguyn tc chung v bioprospecting.
9. Cng ngh nano sinh hc (bionanotechnology)

Nm 1981, cng vi hai nh khoa hc khc, gio s Gerd Binnig (c) c trao gii thng Nobel Vt l v pht minh ra loi knh hin vi mi khng hot ng bng nguyn tc quang hc, m c th tip xc trc tip vi cc nguyn t bng mt u kim nh xu. Vi pht minh ny, GS Binnig m ra cnh ca dn loi ngi n mt cng ngh siu nh, cn gi l cng ngh nano, m sn phm ca n c o bng nanometer (phn triu mm). Ngy nay knh hin vi khng nhn nguyn t na m dng u kim nh li ti tip xc trc tip v iu khin cc nguyn t, di chuyn lp rp chng vi nhau v nh vy c th to nn bt k mt cu trc no. Mc tiu lu di ca cc nh khoa hc t ra cho cng ngh nano l lm ra cc sn phm theo cch cc sinh vt c t nhin sn sinh ra. Trong khi nn cng nghip hin nay ca chng ta lm ra cc sn phm bng cch ca, c, o, phay tin, th trong t nhin cc sinh vt c hnh thnh theo chu trnh nguyn t-phn t-protein-t bo-c th sinh vt. Trong vng 50 nm na cng ngh siu nh nano s em li cho loi ngi nhiu thay i hn tt c nhng thay i con ngi tng tri qua
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t thi trung c n nay. Trong tng lai cng ngh nano s cho php chng ta np ln mt din tch rng bng mt chic mng tay lng thng tin 10.000 gigabyte tng ng vi khong 15.000 a CD. Ngy nay, cc chi tit siu nh c mt trong rt nhiu cc sn phm cng ngh cao, trong trc tin phi k n cc chp in t t hon. Chng ta c th tm thy cc sn phm ca cng ngh siu nh, v d nh trong h thng phun nh xu trong my in phun, c iu khin bng in t phun ra nhng git mc li ti dng bi ln giy, h thng quang hc cn gi l mt thn c thng tin trong my CD, b phn c tinh xo trong chic ng h siu mng ca Thy S hoc u c siu nh trong cng ca mt my vi tnh. Cng ngh nano sinh hc l mt lnh vc trong cng ngh nano lnh vc a ngnh trong khai thc vt liu, thit b hoc sn xut cc cht phm vi kch thc ti hn nm gia chiu di phn t v bc sng nh sng kh kin t 0,1 n 500 nm. Cng ngh nano sinh hc l phng hng mi cho php thu nhn nhng thng tin v h thng sinh hc mc chm lng t, u d kch thc nano ti kch thc mt phn t ring r dng trong chn on bnh, l phng php in stitu mi cung cp thng tin tt hn v chc nng t bo, l cng ngh thao tc ci bin 2 chiu v 3 chiu i vi m v t bo v vn chuyn, phn phi thuc hoc gene vo m v t bo thng qua khng ch kch thc ht, hot ha v gii phng hot cht thuc thng qua c ch v thit b nh bm kch thc nano, van t bo v c quan nhn to. Trong y hc, ngi ta hi vng phu thut gene, phu thut t bo, iu tr t bo, iu tr gene, tng hp gene, chn on ti t bo, cc h thng c kh in t nano y hc cn gi l cc cng c nano thng minh, robot m kch thc nano s c a ra thc hnh trong vng 20 nm sp ti. Tuy nhin, hin nay cng c mt s ng dng ca cng ngh siu nh nano trong y hc. V d nh: * Mt chic my bonh xu bng u mt que dim c a vo c th ngi, lun lch trong mch mu v no vt lp cn bm trn thnh mch. Chic my bo ny chuyn ng nh mt ng c chy bng tuc binh nc c ng knh 2,5 mm. Tt c chic my k diu ny c t gn trn u mt ng si tim mch chuyn phc v cho cc ca m ni soi. * Ko phu thut t si hp kim titan v nickel mng 0,63 mm, c s dng trong ca phu thut thn kinh. Khi thao tc, bc s phu thut khng iu khin ko trc tip bng tay m thng qua mt h thng cc nt cm ng.
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* Mt chp in t sinh hc trn mt lp silic rng chng vi mm2 c th cha ng c hng chc vn phn t mang nhng thng tin v gene. Chp sinh hc l mt dng c ti u phn tch nhanh cc mu mu v m nhanh chng chn on cc khi u c tnh ngay t khi trong giai on sm nht. * Videocamera siu nh kch thc 1130 mm. Vin thuc camera ny c bnh nhn nut vo bng v s cung cp cho bc s nhng hnh nh c th v chnh xc v tnh trng b my tiu ha ca bnh nhn. * Bng cng ngh siu nh nano, cc nh khoa hc c th ch to cc robot nano chuyn truy tm cc t bo ung th tim n trong c th ngi tiu dit chng. Cng ngh nano sinh hc tp trung cho cc nghin cu s dng phn t DNA dng chp lm phng tin lu gi thng tin, to khung 2-3 chiu h tr sinh trng v lin so ca m v t bo cy ghp, cung cp thuc n tn im m v t bo trong iu tr ung th. Cng c nano kh cht c nhim cng nghip trong t, vt liu composit nano ni xng, dy dn bng protein hay DNA trong ng c nano. Cng tc DNA iu khin mch nano, bn nano trong nghin cu hu gii m b gene, xng nano ch to phn t protein. Trong lnh vc mi trng cc loi ht nano va c th lm u d va x l nhim ngun nc, theo di v bo ng v v kh ha hc v sinh hc mt cch l tng v hiu qu. i vi Vit Nam, trong iu kin thc t v i ng v c s vt cht ca mnh, chng ta khng th tin hnh c tt c nhng nh hng chung ca th gii. Tuy nhin, trong hon cnh c th ca tng vng, c bit l thnh ph H Ni, thnh ph H Ch Minh v vin La ng bng sng Cu Long, ty theo nhu cu thc t ca a phng mnh c nhng hng nghin cu c th phc v cho nhu cu kinh t x hi ca khu vc v ca c nc. Nhiu ni cng c th nghin cu ng dng mt s mt sau: * Cng ngh gene: nghin cu s thay i genome ca ngi v cc sinh vt khc di tc ng ca mi trng, ca chin tranh ha hc. Nghin cu c im genome ca cc tc ngi Vit Nam, trc mt l cc genome ti th, NST gii tnh, gp phn vo vic nghin cu y t d phng v iu tr bng liu php gene. M rng s hp tc gia ngnh sinh hc v cc ngnh khc gp phn hin i ha nhng nghin cu v phn loi v nh gi ti nguyn sinh vt. a cng ngh gene vo vic nghin cu sn xut vaccine th h mi v sn xut cc kt chn on bnh
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ngi, ng vt v thc vt. Tng bc a k thut DNA array vo nghin cu v ng dng. * Cng ngh enzyme protein: ng dng enzyme cng nghip vo vic ch bin sn phm nng lm ng nghip v lm thuc cha bnh. ng thi, nhanh chng vic nghin cu proteomics thnh cng c c lc cho vic nh gi chc nng gene v tm kim nhng protein c gi tr dc phm cao, sn xut protein ti t hp v chn on bnh ngi v gia sc. Min dch hc phn t l mt ni dung nghin cu s mang li hiu qu ng dng cao trong sn xut vaccine v kt chn on, khng sinh, vitamin t cng ngh ln men vi sinh vt v vi sinh vt ti t hp. * Cng ngh vi sinh: - Nghin cu ti nguyn vi sinh vt nh gi tnh a dng ca chng cc h sinh thi. Xy dng bo tng vi sinh vt cp quc gia nh mt n v c t cch php nhn c lp trong giao dch v qun l cc chng ging vi sinh vt. - nh gi v khai thc ti nguyn vi sinh vt thng qua profiling cc hot cht, xy dng c s d liu v ti nguyn vi sinh vt. Nghin cu ln lp v tuyn chn cc ging c hot tnh v sn lung cao. - Nghin cu ng dng v pht trin cng nghip ln men vi sinh vt thnh mt ngnh sn xut cc loi sn phm nh nguyn liu lm thuc, khng sinh, vitamin, acid amin, cc ph gia, cc cht mu thc phm, m phm. * Cng ngh t bo ng vt: nghin cu cng ngh sinh sn cloning; cng ngh t bo gc v c s khoa hc ca phng php tr liu t bo; lp bn gene v cc tnh trng tt vt nui; chuyn gene ng vt. * Cng ngh t bo thc vt: nghin cu hon thin qui m cng nghip v chuyn giao cng ngh nhn ging v tnh cy trng nng lm nghip (cy n qu c sn, cy cng nghip, cy hoa, cy cnh v cy lm nghip bn a v cy lai cao sn); tng bc trin khai s dng ch th phn t v lp bn gene vo cng tc chn ging trn c s cng ngh truyn thng v cng ngh t bo; a cng ngh to cy chuyn gene vo thc tin sn xut; ci tin ging cy trng vt nui, phn bn, thuc tr su sinh hc, kt chn on bnh cy trng v vt nui, bo qun v ch bin cc sn phm nng lm ng nghip tng gi tr s dng v thng mi. * Cng ngh sinh hc y dc, sn phm ca chng c hm lng CNSH cao, cht lng tt (ngh en, nhn sm Ngc Linh, v.v.), nhng cht c hot tnh sinh hc mnh ng vt, thc vt v vi sinh vt. Cc vaccine th h mi bao gm vaccine t bo, vaccine ti t hp, vaccine
229 Trng Vn Lung
DNA. Cc loi thuc iu tr c ngun gc protein ti t hp nh khng th a dng v c bit l khng th n dng, cc loi hormone dng protein, cc protein c tr. Cc liu php cng ngh cao nh liu php gene, liu php t bo dng cha nhng bnh him ngho. Trong thi gian ti CNSH s ch trng vo vic phc v lnh vc y dc v an ninh quc phng. Cn hp tc gia cc trng i hc v cc vin nghin cu nghin cu vaccine ti t hp, di truyn phn t ngi v kt chn on bnh truyn nhim v bnh him ngho. Trong thi gian ti cn phi hp vi b Quc phng v b Cng an xc nh hi ct lit s . * Cng ngh sinh hc phc v mi trng l nng cao cht lng nghin cu khoa hc trong lm sch nc thi, kh nitrogen, lm sch nhim du v kim loi nng, cc phng php ty cc cht c ha hc, xy dng c s vt cht trang thit b, o to nhn lc nhm trin khai cc nghin cu cht lng cao phc v an ninh quc phng v dn sinh. * Cng nghip sinh hc (bioindustry). Nhng lnh vc sn xut chnh l:Cng nghip sinh hc y dc (biomidicine) c cc nhm sn phm nh hormone, thuc chng ung th, khng sinh, thuc sinh trng, thuc min dch (vaccine) . Cng ngh sinh hc ha cht (biochemicals) c cc loi polymer sinh hc, acid amin, acid hu c, enzyme cng nghip, cht mu, hot cht b mt. Cng nghip sinh hc mi trng (bioenvironmental) bao gm ch phm vi sinh vt dng lm sch mi trng, kh sulphate, kh thi, kh trng v cht kt dnh. Cng nghip sinh hc thc phm (biofood). Cng nghip sinh hc nng lng v ti nguyn (bioenergy and resources) bao gm kh methan sinh hc, ng lnh nh CO2, sinh khi quang hp, kh sinh hc, bt git vi sinh. Cng nghip sinh hc nng nghip v thy sn (bioagriculture and ocean) tp trung cho ging lai, vaccine th y, sinh phm chn on, phn bn vi sinh, ti nguyn sinh hc bin, nh my thc n chn nui. Cng nghip sinh hc ch bin (bioprocessing) v k ngh sinh hc (bioengineering) bao gm qui trnh cng ngh ln men, k thut nui cy t bo ng vt, k thut nui cy t bo thc vt nhm tng h s nhn ging nhng cy c sn, bn a, nhng cy c gi tr kinh t cao, phc v cho sn xut v i sng. Cng ngh sinh hc kim nh (bioevaluation and verification systems) bao gm cng ngh nh gi hiu qu v bn vng, biosensor, biochip, cng ngh chn on, thit b kim nh sinh hc. R rng, CNSH l ci cha kha m ng cho s pht trin nn kinh t ca t nc. Cuc cch mng khoa hc k thut ni chung v cuc cch mng CNSH ni ring thu ht nhiu ngi trn tri t ny tham gia vo s nghip cao c .
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TI LIU THAM KHO
1. L Trn Bnh, 2003. nh hng nghin cu v trin khai ca vin

Cng ngh sinh hc. Bo co khoa hc Hi ngh Cng ngh sinh hc ton quc 16-17 thng 12 nm 2003. Nxb Khoa hc v K thut H Ni, tr: 48-51 L Trn Bnh, 2004. Nhng thnh tu ni bt trong nghin cu c bn ca khoa hc s sng trong 10 nm qua. Bo co khoa hc Hi ngh ton quc Nhng vn nghin cu c bn trong khoa hc s sng Nxb Khoa hc v K thut H Ni, tr: 17-22. Phm Hu Gic, 1999. nh hng pht trin cng ngh sinh hc Vit Nam n 2010. Bo co khoa hc Hi ngh Cng ngh sinh hc ton quc 9-10 thng 12 nm 1999. Nxb Khoa hc v K thut H Ni, tr: 37-46. Phm Hu Gic, L Minh St, 2003. Cc chnh sch v nh hng pht trin cng ngh sinh hc thi gian ti Vit Nam. Bo co khoa hc Hi ngh Cng ngh sinh hc ton quc 16-17 thng 12 nm 2003. Nxb Khoa hc v K thut H Ni, tr: 44-47. Nguyn Ngc Knh, 1999. Chng trnh k thut kinh t v cng ngh sinh hc n nm 2010.Bo co khoa hc Hi ngh Cng ngh sinh hc ton quc 9-10 thng 12 nm 1999. Nxb Khoa hc v K thut H Ni, tr: 47-51. Trn Duy Qu, 2003. Chng trnh nghin cu v pht trin cng ngh sinh hc: thnh tu v thch thc. Bo co khoa hc Hi ngh Cng ngh sinh hc ton quc 16-17 thng 12 nm 2003. Nxb Khoa hc v K thut H Ni, tr: 39-43. L Vn, 2004. Ngn hng t bo gc u tin trn th gii, Tp ch Th gii mi, s 595, tr: 56-59. Nng Vnh, 1999. Mt s thnh tu v hng pht trin ca cng ngh t bo thc vt c trin vng ng dng nc ta. Bo co khoa hc Hi ngh Cng ngh sinh hc ton quc 9-10 thng 12 nm 1999. Nxb Khoa hc v K thut H Ni, tr: 62-70.
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Trao i trc tuyn ti:

1 CNSH-M u Lung Trng Vn

2 CNSH-M u Lung Trng Vn

2. Lch s pht trin CNSH

3 CNSH-M u Lung Trng Vn

3. Ha hn ca CNSH vi cc nc ang pht trin

4 CNSH-M u Lung Trng Vn

5 CNSH-M u Lung Trng Vn

6 CNSH-M u Lung Trng Vn

7 CNSH-M u Lung Trng Vn

8 CNSH-M u Lung Trng Vn

Cng ngh sinh hc vi nng lng

Anode Cathode Mng Pin sinh hc n gin H2 O2

Ngi ta cng c th dng enzyme bin i mt s cht to H2.

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Cng ngh sinh hc phc v nng lm ng nghip

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c gn vo b gene ca thc vt s lm xut hin khi u hoc r tc v chng cha oncogene.

Hnh II.2. M hnh plasmid Ti

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Thu nhn phi

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Sinh thit phi

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