Prokaryotic Expression and Protein Purification of O-type Foot-and-mouth Disease Virus VP1 Gene

With O-FMDV recombinant plasmid pMD18T-O-VP1 as a template,O-FMDV-VP1 gene fragments were obtained by using PCR amplification.This gene fragments wwas connected with prokaryotic expression vector pET32a to construct recombinant expression vector pET32a-O-VP1.After PCR identification and sequencing,IPTG was used to induce the expression of VP1 gene.Different induction time of the bacterium were collected for SDS-PAGE electrophoresis.The best induction time was mastered.And the electrophoresis film with the best induction time was cut to do Western-blotting.The expression product testing with its anti-serum responsiveness was analyzed.The results showed that the protein with the molecular weight of about 45 ku had significant strip reaction and it could be identified by positive serum of food-and-mouth disease.These results showed that the FMDV-VP1 genes were efficiently expressed in E.coli.The purified expression protein is expected to develop diagnostic antigen and peptide vaccines.