Use of bacteriophage lambda recombination functions to promote gene replacement in Escherichia coli
Department of Molecular Genetics and Microbiology
Bacteriophage lambda; Calcium; Chromosomes, Bacterial; Cloning, Molecular; DNA Primers; DNA, Superhelical; Escherichia coli; Gene Targeting; Genotype; Operon; Plasmids; Polymerase Chain Reaction; Recombinant Fusion Proteins; *Recombination, Genetic; Restriction Mapping
Life Sciences | Medicine and Health Sciences
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.
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Citation: J Bacteriol. 1998 Apr;180(8):2063-71.
Journal of bacteriology
Murphy, Kenan C., "Use of bacteriophage lambda recombination functions to promote gene replacement in Escherichia coli" (1998). Open Access Articles. 660.