Академический Документы
Профессиональный Документы
Культура Документы
Dr.T.V.Rao MD
Dr.T.V.Rao MD
Dr.T.V.Rao MD
Dr.T.V.Rao MD
Conjugation
Dr.T.V.Rao MD
Simple Conjugation
Dr.T.V.Rao MD
Conjugation
Dr.T.V.Rao MD
F factor
Transfer factor that contains the genetic information necessary for synthesis of Sex Pilus and for self transfer without any other identifiable genetic materials such as drug resistance
Dr.T.V.Rao MD 10
T.V.Rao MD
Can transform chromosomal genes to recruitment with high frequency are known as Hfr cells
Conversion of F+ cells into Hfr state is reversible. F factor incorporates some chromosomal genes and is called as F Sexduction The process of transfer of host genes Dr.T.V.Rao MD through F factor
11
Dr.T.V.Rao MD
13
RTF
Shigella + E.coli excreted in the stool resistant to several drugs in vivo and vitro Plasmid mediated transmitted by Conjugation Episomes spread the resistance
Dr.T.V.Rao MD 16
Dr.T.V.Rao MD
17
Hfr Conjugation
Dr.T.V.Rao MD
18
Dr.T.V.Rao MD
19
Dr.T.V.Rao MD
20
Composition of RTF
Plasmid consists of two components A transfer factor RT, helps conjugational transfer and resistant determinants ( r ) to each of the several drugs RTF + r determinants are known as R factor
Dr.T.V.Rao MD 21
R factor
R factor can contain several determinants as many as 8 or > 8 drugs Guide the cell for production of Enterotoxins too But R factors can be inhibited by Bile salts R factors can be transferred to animals
Dr.T.V.Rao MD 22
Genesis of R factors
In discriminate use of Antibiotics in vet nary Medicine has increased the spread of R factors to Human Addition of Antibiotics to Animal feeds to be prohibited.
Dr.T.V.Rao MD 23
Dr.T.V.Rao MD
25
Mutations
Mutilations can be 1 Stepwise mutation as in Penicillin use 2 One step mutation Streptomycin use May show low resistance or High resistance If tuberculosis is treated with sole drug as of Only Streptomycin some resistant mutants appear and replaces sensitive bacteria in due course so the occurrence of MDR - TB
Dr.T.V.Rao MD 26
Other Mechanisms
Use of Penicillin created resistant Staphylococcus by transduction R factors created resistance to several drugs, caused increased virulence Spread to several humans and animals
Molecular Genetics
Analysis and manipulation of DNA using Biochemical and Microbiological techniques
Dr.T.V.Rao MD 30
Genetic Engineering
Under standing Molecular genetics in Biochemistry fuels genetic Engineering Recombinant DNA (renal) techniques changed the ideals of Medicine
Genetic Engineering
Genetic Engineering Was Born from
Genetic Recombination
Genetic engineering involves changing the genetic material in an organism to alter its traits or products A recombinant DNA molecule contains DNA fragments spliced together from 2 or more organisms
Dr.T.V.Rao MD 32
Modern applications
Pharmaceutical production
Insulin, interferon, hormones, vaccines etc.
Genetically engineered plants Animal gene alterations Gene probes DNA fingerprinting The human genome initiative
Dr.T.V.Rao MD 33
Genetic Engineering
Isolation of Genes coding for any desired protein from Microorganism or from cell of higher life forms including human beings and their introduction into a suitable microorganism in which genes would function directing the production of specific proteins
Dr.T.V.Rao MD 34
Dr.T.V.Rao MD
35
Dr.T.V.Rao MD
36
Hepatitis B vaccine
Dr.T.V.Rao MD
37
Restriction Endonucleases
A restriction enzyme (or restriction endonuclease) is an enzyme that cuts double-stranded DNA. The enzyme makes two incisions, one through each of the sugar-phosphate backbones (i.e., each strand) of the double helix without damaging the nitrogenous bases They work with cutting up foreign DNA, a process called
Dr.T.V.Rao MD 38
Dr.T.V.Rao MD
39
DNA Probes
There are Radioactive Biotinylated otherwise labeled copies united single stranded DNA Contains 20 -25 nucleotides Helps detection of Homology DNA by Hybridization. Helps Diagnosis of Infectious Diseases Minute quantities of DNA can be detected.
Dr.T.V.Rao MD 40
Blotting Techniques
Drug fragments obtained by restriction enzyme digestion on separation Gel can be transferred to Nitrocellulose or nylon membranes Several methods 1 Southern blotting 2 Northern Blotting 3 Western blotting
Dr.T.V.Rao MD 41
western blot
The western blot (alternatively, protein immunoblot) is an analytical technique used to detect specific proteins in a given sample of tissue homogenate or extract. It uses gel electrophoresis to separate native or denatured proteins by the length of the polypeptide (denaturing conditions) or by the 3-D structure of the protein (native/ non-denaturing conditions)
Dr.T.V.Rao MD 42
Western Blotting
In Western Blot Protein ( Antigen ) mixture is separated by SDS ( Sodium dodecyl sulfate polyacrylamide gel electrophoresis ) Blotted on to Nitro cellulose strips and identified by radio labeled or enzyme labeled antibodies as probes
Dr.T.V.Rao MD 43
Western Blot
Dr.T.V.Rao MD
44
Western Blot testing is confirmatory test for diagnosis of HIV/AIDS Identifies antibodies directed against different antigens in pathogen Surface, Core Dr.T.V.Rao MD RT antigen
Western Blot to confirm HIV Infections made land mark Diagnostic tool
45
Dr.T.V.Rao MD
46
PCR -Sequences
PCR consists of several cycles of sequential DNA replication where the products of first cycle becomes the template for the Next It makes available abundant quantities of specific DNA sequences starting
Dr.T.V.Rao MD 49
Genetic Mapping
Genetic sequences for Bacteriophages and virus Genetic mapping is done most of the Human Genes
Dr.T.V.Rao MD
50
Dr.T.V.Rao MD
51
Dr.T.V.Rao MD
52
Dr.T.V.Rao MD
54
Dr.T.V.Rao MD
55
Dr.T.V.Rao MD
56
The Programme Created by Dr.T.V.Rao MD for Medical Students in the Developing World
Email
doctortvrao@gmail.com
Dr.T.V.Rao MD
57