title: Isolation of alpha-toxin, theta-toxin and kappa-toxin mutants of Clostridium perfringens by Tn916 mutagenesis.
abstract: Clostridium perfringens is the causative agent of clostridial myonecrosis or gas gangrene and mediates infection and disease by producing numerous extracellular toxins, including alpha-toxin, theta-toxin and kappa-toxin. Tn916-mutagenesis was used to isolate mutants defective in their ability to produce either alpha-toxin or theta-toxin. Nine independently derived mutants were isolated. In four of these mutants Tn916 had inserted at sites located 193 bp or 198 bp upstream of the theta-toxin structural gene, pfoA. Four mutants contained large deletions, three in regions which encompassed the theta-toxin structural and regulatory genes pfoA and pfoR, respectively, and the kappa-toxin structural gene, colA, and one in a region encompassing the alpha-toxin structural gene, plc. These mutants should prove to be invaluable for further genetic studies aimed at determining the role of these toxins in virulence.
authors.affiliation: Department of Microbiology, Monash University, Clayton, Victoria, Australia.
authors.forename: M M
authors.initials: MM
authors.lastname: Awad
authors.forename: J I
authors.initials: JI
authors.lastname: Rood
mesh_terms: D001427:Bacterial Toxins / Q000235:genetics*; D001483:Base Sequence; D002135:Calcium-Binding Proteins; D002874:Chromosome Mapping; D003016:Clostridium perfringens / Q000235:genetics*; D017364:Collagenases / Q000235:genetics*; D004251:DNA Transposable Elements; D004269:DNA, Bacterial; D016521:Electrophoresis, Gel, Pulsed-Field; D013293:Enterococcus faecalis / Q000235:genetics; D006460:Hemolysin Proteins; D008969:Molecular Sequence Data; D016254:Mutagenesis, Insertional; D016133:Polymerase Chain Reaction; D010738:Type C Phospholipases
chemical_list: D001427:Bacterial Toxins; D002135:Calcium-Binding Proteins; D004251:DNA Transposable Elements; D004269:DNA, Bacterial; D006460:Hemolysin Proteins; C020452:Clostridium perfringens theta-toxin; D010738:Type C Phospholipases; C040649:alpha toxin, Clostridium perfringens; D017364:Collagenases