AMPLIFICATION AND IDENTIFICATION OF ALPHA TOXIN GENE IN Clostridium perfringens TYPE A

To detect Clostridium Perfringens type A exactly and quickly,the biochemical characteristics of CVCC38 Clostridium Perfringens type A were determined by culturing in anaerobic meat liver soup,Gram's stain and culturing in iron milk medium in this study.Alpha toxin protein was roughly extracted by using ammonium sulfate(saturation for 60%) to dialysis purify and identify it by SDS-PAGE.Bioactivity of the purified alpha toxin was detected by injected into mouse peritoneal cavity.The results showed that anaerobic meat liver soup was turned turbid and produced large amounts of gas;the iron milk medium produced blasting fierce fermentation;alpha toxin molecular weight was 43.6kDa;the mouse appeared clinical symptoms of clostridial disease.This study made important experimental basis for identification and extraction of alpha toxin to explore the pathogenesis mechanism of it and prepare the toxoid vaccine.