Construction of an alpha toxin gene knockout mutant of Clostridium perfringens type A by use of a mobile group II intron

Appl Environ Microbiol. 2005 Nov;71(11):7542-7. doi: 10.1128/AEM.71.11.7542-7547.2005.


In developing Clostridium perfringens as a safe vaccine vector, the alpha toxin gene (plc) in the bacterial chromosome must be permanently inactivated. Disrupting genes in C. perfringens by traditional mutagenesis methods is very difficult. Therefore, we developed a new strategy using group II intron-based Target-Tron technology to inactivate the plc gene in C. perfringens ATCC 3624. Western blot analysis showed no production of alpha toxin protein in the culture supernatant of the plc mutant. Advantages of this technology, such as site specificity, relatively high frequency of insertion, and introduction of no antibiotic resistance genes into the chromosome, could facilitate construction of other C. perfringens mutants.

Publication types

  • Evaluation Study
  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Clostridium perfringens / genetics*
  • Gene Deletion*
  • Gene Targeting / methods
  • Genetic Techniques
  • Introns / genetics*
  • Mutagenesis, Insertional / methods*
  • Promoter Regions, Genetic
  • Type C Phospholipases / genetics*


  • Type C Phospholipases