Use of bacteriophage lambda recombination functions to promote gene replacement in Escherichia coli.
Replacement of Escherichia coli's RecBCD function with phage lambda's Red function generates a strain whose chromosome recombines with short linear DNA fragments at a greatly elevated rate. The rate is at least 70-fold higher than that exhibited by a recBC sbcBC or recD strain. The value of the system is highlighted by gene replacement with a PCR-generated DNA fragment. The deltarecBCD::Plac-red kan replacement allele can be P1 transduced to other E. coli strains, making the hyper-Rec phenotype easily transferable.
Murphy
K C
KC
Department of Molecular Genetics and Microbiology, University of Massachusetts Medical School, Worcester 01655, USA. kenan.murphy@ummed.edu
D010582:Bacteriophage lambda / Q000235:genetics*; D002118:Calcium / Q000494:pharmacology; D002876:Chromosomes, Bacterial / Q000235:genetics*; D003001:Cloning, Molecular / Q000379:methods; D017931:DNA Primers; D004278:DNA, Superhelical; D004926:Escherichia coli / Q000187:drug effects / Q000235:genetics* / Q000821:virology; D018390:Gene Targeting / Q000379:methods*; D005838:Genotype; D009876:Operon; D010957:Plasmids; D016133:Polymerase Chain Reaction; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis; D011995:Recombination, Genetic; D015183:Restriction Mapping
D017931:DNA Primers; D004278:DNA, Superhelical; D011993:Recombinant Fusion Proteins; D002118:Calcium