Abstract:
:Recombineering is an efficient method of in vivo genetic engineering applicable to chromosomal as well as episomal replicons in Escherichia coli. This method circumvents the need for most standard in vitro cloning techniques. Recombineering allows construction of DNA molecules with precise junctions without constraints being imposed by restriction enzyme site location. Bacteriophage homologous recombination proteins catalyze these recombineering reactions using double- and single-stranded linear DNA substrates, so-called targeting constructs, introduced by electroporation. Gene knockouts, deletions and point mutations are readily made, gene tags can be inserted and regions of bacterial artificial chromosomes or the E. coli genome can be subcloned by gene retrieval using recombineering. Most of these constructs can be made within about 1 week's time.
journal_name
Nat Protocjournal_title
Nature protocolsauthors
Sharan SK,Thomason LC,Kuznetsov SG,Court DLdoi
10.1038/nprot.2008.227subject
Has Abstractpub_date
2009-01-01 00:00:00pages
206-23issue
2eissn
1754-2189issn
1750-2799pii
nprot.2008.227journal_volume
4pub_type
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