Cloning, expressing and purification of recombinan

Abstract

ToxA has been documented to be the main toxin of Pasteurella multocida. Therefore, it is a potential factor for the studies to produce recombinant vaccines against Pasteurellosis in pigs. In this study, we cloned and expressed Tox1 protein, a N-fragment of ToxA (from amino acid 1 to amino acid 487), in Escherichia coli BL21 with pET32b vector. The recombinant Tox1 protein was expressed well when induced with 1mM IPTG and 3% ethanol. The protein was denaturated and dissolved in lysis buffer containing 6M urea and purified via Ni-NTA column. The SDS-PAGE and Western blot results showed that Tox1 was correctly expressed and well-purified. This is the preliminary material for our further studies to produce recombinant vaccine against Pasteurellosis in pigs caused by Pasteurella multocida