Detection, cloning, and sequencing of the enterotoxin gene of Clostridium perfringens type C isolated from goat

Clostridium perfringens enterotoxin (CPE) is an important virulence factor involved in C. perfringens food poisoning and other gastrointestinal illnesses. In our experiment a Clostridium perfringens enterotoxin gene (cpe) positive type C strain was obtained by multiplex PCR from 34 C. perfringens isolates from goat in Guizhou Province, China. Th en a pair of specifi c primers was designed and synthesized according to the published cpe gene, and the complete cpe gene was amplifi ed from the genomic DNA of the cpe-positive C. perfringens isolate. Th e PCR product was cloned into pMD18-T vectors and transformed into competent cell DH5a. Th e recombinant plasmid was identifi ed by PCR and restriction enzyme then sequenced and analyzed. Th e results showed that the cloned gene was 960 bp in length; the gene coded 319 amino acids, which shared 99.4~99.8% sequence identity and 99.1~-99.7% amino acid sequence identity with reference strains in GenBank. Th ese results provide a basis for further research on the structure, properties, and biological activities of Clostridium perfringens enterotoxin. It also set the groundwork for further investigation of the mechanism of disease caused by C. perfringens enterotoxin.