Site-Specific Recombination in E. coli
Explore the mechanism of site-specific recombination in E. coli, including details on attachment sites, bacteriophage recombination, advantages of site-directed recombination cloning, and more. Discover the integration process, structures of attB, attP, attR, and attL, as well as the potential benefits of this technique over traditional cloning methods.
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Presentation Transcript
Site-specific recombination of into E. coli integrates at one specific site in the E. coli chromosome The site is, located between the gal and bio operons called the attachment site designated attB (attachment Bacterial). 30 bp long contains a conserved central 15 bp for recombination. usually represented as BOB', where B and B' represent the bacterial DNA on either side of the conserved central element.
The bacteriophage recombination site - attP more complex contains the identical central 15 bp region as attB. overall structure can be represented as POP'.
The result of recombination integrated prophage is flanked by two slightly different attachment sites attR has the structure POB attL has the structure BOP'
Potential advantages with site-directed recombination cloning higher efficiency and accuracy than ligation-based cloning shorter training time of personnel can process multiple samples in parallel less need for confirmation after cloning does not require restriction endonuclease digest of vector or insert before insertion of fragment no need for curing vector or insert of restriction sites can clone in large vectors, or with large inserts
