Targeted allele replacement mutagenesis of Corynebacterium pseudotuberculosis.

ABSTRACT: A two-step allele replacement mutagenesis procedure, using a conditionally replicating plasmid, was developed to allow the creation of targeted, marker-free mutations in Corynebacterium pseudotuberculosis. The relationship between homologous sequence length and recombination frequency was determined, and enhanced plasmid excision was observed due to the rolling-circle replication of the mutagenesis vector. Furthermore, an antibiotic enrichment procedure was applied to improve the recovery of mutants. Subsequently, as proof of concept, a marker-free, cp40-deficient mutant of C. pseudotuberculosis was constructed.

SUBMITTER: Walker CA 

PROVIDER: S-EPMC3126432 | biostudies-literature | 2011 May

REPOSITORIES: biostudies-literature