Targeted allele replacement mutagenesis of Corynebacterium pseudotuberculosis

Caray A. Walker, Willie Donachie, David George Emslie Smith, Michael C. Fontaine

    Research output: Contribution to journalArticle

    2 Citations (Scopus)

    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.

    Original languageEnglish
    Pages (from-to)3532-3535
    Number of pages4
    JournalApplied and Environmental Microbiology
    Volume77
    Issue number10
    DOIs
    Publication statusPublished - May 2011

    Keywords

    • Alleles
    • Anti-Bacterial Agents
    • Corynebacterium pseudotuberculosis
    • DNA, Bacterial
    • Gene Deletion
    • Gene Targeting
    • Genetic Vectors
    • Molecular Sequence Data
    • Mutagenesis, Insertional
    • Plasmids
    • Selection, Genetic
    • Sequence Analysis, DNA

    Fingerprint Dive into the research topics of 'Targeted allele replacement mutagenesis of <i>Corynebacterium pseudotuberculosis</i>'. Together they form a unique fingerprint.

  • Cite this