Cloning of Tpel Gene of Clostridium perfringens in E. coli

Clostridium perfrinjens is an anaerobics, Gram-positive, rod-shaped and heat resistant bacterium of genus clostridium. C. perfringens is a spore-forming bacterium and widely occurring pathogen. The organism is grouped into 5 types (A, B, C, D, and E) on the basis of the production of 4 major toxins alpha, beta, epsilon, and iota toxins. Tpel Clostridium perfringens (C. perfringens) toxin have identified with A, B, and C types by cytotoxin activity in recent years. In this study C. perfringens type B had been used. Tpel caused to intestinal disease especially intestinal infections in human and necrotic enteritis in birds. In this study, perfect genomic DNA extracted by phenol-chloroform and Polymerase Chain Reaction (PCR) method used to isolation Tpel gene by a couple exclusive primer of perfect bacterium genomic DNA. PCR product after joining to pTZ57RL/T vector by TA cloning method in E. coli strain TOP10 susceptible became cloned and then colony PCR method used to screening transforming bacterium colonies with recombinant plasmid. Presence of fragment close to 2469bp on 1% agarose gel indicated that Tpel gene in E. coli strain TOP10 have be cloned.