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.