Expression of TDH from Vibrio parahaemolyticus and Preparation of Monoclonal Antibody against TDH

Abstract:

To express of tdh gene in E.coli，TCBS medium was used to select the suspected strains of Vibrio parahaemolyticus （VP）. Primers were designed and synthesized to identify VP and amplify tdh gene based on the gene sequence from GenBank. The PCR gene product was ligated into prokaryotic expression vector pET-28a，and then the recombinant plasmids pET-28a-tdh was transformed into E.coli Rosetta. TDH protein was expressed by IPTG induction. To prepare monoclonal antibody against TDH （thermostable direct hemolysin） produced by VP，the purified TDH was used to immune BALB/c mice. The hybrid tumor cells named as T9N10 were prepared to secrete monoclonal antibodies by conventional prepared techniques. The secreted monoclonal antibodies are IgG1，which was 146 000 weight，and showed strong specificity. Results provide new insights to immunological rapid detection and research.