Abstract
Recombineering is a recently developed method of in vivo genetic engineering used in Escherichia coli and other Gram-negative bacteria. Recombineering can be used to create single-base changes, small and large deletions, and small insertions in phage lambda as well as in bacterial chromosomes, plasmids, and bacterial artificial chromosomes (BACS). This technique uses the bacteriophage lambda generalized recombination system, Red, to catalyze homologous recombination between linear DNA and a replicon using short homologies of 50 base pairs. With recombineering, single-stranded oligonucleotides or double-stranded PCR products can be used to directly modify the phage lambda genome in vivo. It may also be possible to modify the genomes of other bacteriophages with recombineering.
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