Electroporation of DNA sequences from the pathogenicity locus (PaLoc) of toxigenic Clostridium difficile into a non-toxigenic strain

G. Ackermann, Y. J. Tang, J. P. Henderson, A. C. Rodloff, J. Silva, Stuart H Cohen

Research output: Contribution to journalArticle

11 Scopus citations

Abstract

Toxigenic Clostridium difficile is the etiologic agent of C. difficile-associated diarrhoea (CDAD), the most common cause of hospital-acquired infectious diarrhoea. The genes tcdA and tcdB, which encode for the toxin A and B proteins, are part of the pathogenicity locus (PaLoc) of toxigenic C. difficile. Genetic and virulence studies at the molecular level in C. difficile have been hindered by the lack of techniques for DNA manipulation in this species. We describe the electroporation of DNA fragments from a toxigenic isolate into a non-toxigenic strain of C. difficile. Using previously described methods of electroporation into Clostridium spp., the complete toxin B gene and polymerase chain reaction (PCR) fragments of the PaLoc were cloned and electroporated into a non-toxigenic strain of C. difficile. The resulting transformed clones were screened for the introduced gene fragments by PCR, which confirmed their presence. This is the first description of introduction of DNA into C. difficile by electroporation.

Original languageEnglish (US)
Pages (from-to)301-306
Number of pages6
JournalMolecular and Cellular Probes
Volume15
Issue number5
DOIs
StatePublished - 2001

Keywords

  • Clostridium difficile
  • Electroporation

ASJC Scopus subject areas

  • Cell Biology
  • Molecular Biology

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