CLONING AND EXPRESSION MOST EXPECTED ANTIGENIC FRAGMENT OF BETA-TOXIN GENE FROM CLOSTRIDIUM PERFRINGENS TYPE B

Abstract

Clostridium perfringens type B and C is an important pathogen and produces Beta-toxin which are responsible necrotic enteritis in humans or livestock. The death in individuals with this disease are over 50%. Vaccines against C. perfringens type B and C are currently manufactured using Beta-toxin produced by the virulent C. perfringens strain itself. To achieve the effective components for the creation of immunity at the first step used different primers in various location of Beta-toxin gene (cbp) by bioinformatics tools according to the secondary protein structure. After amplication of PCR products, one regions of Beta-toxin gene with high antigenicity was cloned into pTZ57RT and sub-cloned into the expression vector pET21a(+). The cloned vector was transformed into E. coli BL21 (DE3) and successfully expressed. Protein expression was confirmed by SDS-PAGE electrophoresis and western blotting. This recombinant peptide from most antigenic region of Beta-toxin gene can be suggested for antibody production and new peptide vaccine.