Use of Bacteriophage λ Recombination Functions To Promote Gene Replacement in Escherichia coli

Replacement of Escherichia coli’s RecBCD function with phage λ’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 ΔrecBCD::P(lac)-red kan replacement allele can be P1 transduced to other E. coli strains, making the hyper-Rec phenotype easily transferable.