1. Endonuklease : memotong DNA dari dalam 2. Eksonuklease : memotong DNA dari ujung Restriction endonuclease Restriction enzymes : DNAase that cleave DNA at specific site Produced by bacteria typically recognize the restriction sites (4- to 8-bp sequences) cleave both DNA strands at this site. commonly are short palindromic sequences the restriction- site sequence is the same on each DNA strand when read in the 5 3 direction.
For each restriction enzyme, bacteria also produce a modification enzyme, which protects a bacteriums own DNA from cleavage by modifying it at or near each potential cleavage site.
The modification enzyme adds a methyl group to one or two bases, usually within the restriction site. Restriction endonuclease Restriction enzymes : DNAase that cleave DNA at specific site When a methyl group is present there, the restriction endonuclease is prevented from cutting the DNA.
Together with the restriction endonuclease, the methylating enzyme forms a restriction-modification system that protects the host DNA while it destroys incoming foreign DNA (e.g., bacteriophage DNA or DNA taken up during transformation) by cleaving it at all the restriction sites in the DNA.
Many restriction enzymes make staggered cuts in the two DNA strands at their recognition site, generating fragments that have a single-stranded tail at both ends. Restriction endonuclease Restriction enzymes : DNAase that cleave DNA at specific site The tails on the fragments generated at a given restriction site are complementary to those on all other fragments generated by the same restriction enzyme.
At room temperature, these single-stranded regions, often called sticky ends, can transiently base-pair with those on other DNA fragments generated with the same restriction enzyme.
A few restriction enzymes, such as AluI and SmaI, cleave both DNA strands at the same point within the restriction site, generating fragments with blunt (flush) ends in which all the nucleotides at the fragment ends are base-paired to nucleotides in the complementary strand. Cleavage of DNA by the restriction enzyme EcoRI This restriction enzyme from E. coli makes staggered cuts at the specific 6-bp inverted repeat (palindromic) sequence shown, yielding fragments with single-stranded, complementary sticky ends.
Many other restriction enzymes also produce fragments with sticky ends. Restriction Enzyme
Source
DNA Sequence Recognized
Ends of Cleaved Molecule
EcoRI
Escherichia coli
5'G AATTC 3'CTTAA G BamHI
Bacillus amyloliquefaciens
5'G GATCC 3'CCTAG G HindIII
Haemophilus influenzae
5'A AGCTT 3'TTCGA A
MstII
Microcoleus species
5'CCTNAGG 3'GGANTCC
TaqI
Thermus aquaticus
5'T CGA 3'AGCT NotI
Nocardia otitidis
5'GCGGCCGC 3'CGCCGGCG AluI*
Arthrobacter luteus
5'AGCT 3'TCGA * = blunt ends
Restriction Enzyme: Source and Site of Action Restriction Cleavage of DNA