Use of the lambda Red recombinase system to rapidly generate mutants in Pseudomonas aeruginosa.

ABSTRACT:

Background

The Red recombinase system of bacteriophage lambda has been used to inactivate chromosomal genes in various bacteria and fungi. The procedure consists of electroporating a polymerase chain reaction (PCR) fragment that was obtained with a 1- or 3-step PCR protocol and that carries an antibiotic cassette flanked by a region homologous to the target locus into a strain that expresses the lambda Red recombination system.

Results

This system has been modified for use in Pseudomonas aeruginosa. Chromosomal DNA deletions of single genes were generated using 3-step PCR products containing flanking regions 400-600 nucleotides (nt) in length that are homologous to the target sequence. A 1-step PCR product with a homologous extension flanking region of only 100 nt was in som

SUBMITTER: Lesic B 

PROVIDER: S-EPMC2287187 | biostudies-literature | 2008 Feb

REPOSITORIES: biostudies-literature