Expression of clostridium difficile toxin B C-terminal repeated unit in E.coli and its immunogenicity / 基础医学与临床

ABSTRACT

Objective To perform expression and to detect immunogenicity of Clostridium difficile Toxin B receptor binding zone(CD3).Methods The clostridium difficile toxin B C-terminal repeated gene was amplified by PCR and cloned into the prokaryotic expression vector pET 22b(+),and the recombinant plasmid pET 22b(+)CD3 was then transformed into E.coli strainBL21(DE3).The E.coli strainBL21(DE3) containing pET22b(+)CD3 was induced with IPTG and analyzed with SDS PAGE.Results A 71.3 ku protein was harvested after inducing with IPTG and thin layer scanning which takes 34% of the total bacterial protein,22.7% of the supernatant and 24.9% of the inclusion body.The contentration of recombinant protein is 0.781 g/L identified by Coomassie brilliant blue G-250 chromatometry method.Conclusion The high expression and purification of clostridium difficile toxin B receptor gene lay a foundation for the further study on CD3 function and clostridium difficile vaccine.