Cloning of VP1 Gene of Foot-and-Mouth Disease (FMDV) Type O and Its Expression in Escherichia coli

In this study, a special primer pair containing BamHⅠand HindⅢaccording to complete genome of foot-and-mouth disease (FMDV) Type O was designed to amplify VP1 gene from plasmid pMD-P1 of FMDV type O. The interest fragment was inserted into PMD18-T vector and the recombinant plasmid was indentifed and the sequence was detected . The target gene VP1 was subcloned into PET32A vector digested by BamHⅠand HindⅢ respectively. Positive clone with interest gene was identified by restriction analysis、PCR and DNA sequencing. Then the recombinant plasmid was transformed into Escherichia coli BL21(DE3) for VP1 expression. The interest gene was induced to express in E.coli with different concentration IPTG. The bacteria containing PET32A-VP1 was collected and examined by SDS-PAGE and western-blotting. Result showed that the structural protein was expressed in E.coli and the amount of expression was higher. Melecular weight of the protein was 41 ku.The protein can be recognized by the positive serum of FMDV, the amount of protein is about 30% of the total bacteria protein. This study showed that the structural protein VP1 amount of expression was higher in E.coli and the protein has antigenicity.