title: Construction of an alpha toxin gene knockout mutant of Clostridium perfringens type A by use of a mobile group II intron.
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.
authors.affiliation: Department of Infectious Diseases and Microbiology, Graduate School of Public Health, University of Pittsburgh, Pittsburgh, PA 15261, USA. cheny@pitt.edu
authors.forename: Yue
authors.initials: Y
authors.lastname: Chen
authors.forename: Bruce A
authors.initials: BA
authors.lastname: McClane
authors.forename: Derek J
authors.initials: DJ
authors.lastname: Fisher
authors.forename: Julian I
authors.initials: JI
authors.lastname: Rood
authors.forename: Phalguni
authors.initials: P
authors.lastname: Gupta
mesh_terms: D003016:Clostridium perfringens / Q000235:genetics*; D017353:Gene Deletion; D018390:Gene Targeting / Q000379:methods; D005821:Genetic Techniques; D007438:Introns / Q000235:genetics*; D016254:Mutagenesis, Insertional / Q000379:methods*; D011401:Promoter Regions, Genetic; D010738:Type C Phospholipases / Q000235:genetics*
chemical_list: D010738:Type C Phospholipases