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

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

Applied and environmental microbiology (2005-11-05)
Yue Chen, Bruce A McClane, Derek J Fisher, Julian I Rood, Phalguni Gupta
ABSTRACT

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.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
TargeTron Vector pJIR750ai, Expression Vector for Bacterial Gene Knockout