Targeted allele replacement mutagenesis of Corynebacterium pseudotuberculosis

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

    Research output: Contribution to journalArticlepeer-review

    2 Citations (Scopus)


    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
    Issue number10
    Publication statusPublished - May 2011


    • 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


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

    Cite this