Targeted allele-replacement mutagenesis of Corynebacterium pseudotuberculosis.

Walker, C. A., Donachie, W., Smith, D.G.E. and Fontaine, M.C. (2011) Targeted allele-replacement mutagenesis of Corynebacterium pseudotuberculosis. Applied and Environmental Microbiology, 77(10), pp. 3532-3535. (doi: 10.1128/AEM.01740-10)

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 recovery of mutants. Subsequently, as proof-of-concept, a marker-free, cp40-deficient mutant of C. pseudotuberculosis was constructed

• Item Type: Articles
• Status: Published
• Refereed: Yes
• Glasgow Author(s) Enlighten ID: Smith, Professor David
• Authors: Walker, C. A., Donachie, W., Smith, D.G.E., and Fontaine, M.C.
• College/School: College of Medical Veterinary and Life Sciences > School of Infection & Immunity
• Journal Name: Applied and Environmental Microbiology
• Publisher: American Society for Microbiology
• ISSN: 0099-2240
• ISSN (Online): 1098-5336