Expression and purification of protective antigen partial genes of vesicular stomatitis virus G protein

According to the published sequence of vesicular stomatitis virus(VSV) G gene in GenBank,a pair of primers was designed and synthesized.The sequence of G gene about 810 bp was amplified by RT-PCR.The amplicon was inserted into pGEX-4T-1 vector,and the recombinant plasmid was identified by PCR amplification and digestion with restriction endonucleases,and then sequenced.The positive recombinant pGEX-G810 was transformed into E.coli BL21(DE3).The expressed recombinant protein was identified by SDS-PAGE and Western blotting.The results showed that,the recombinant protein with a molecular weight of 60ku was soluble in the form of inclusion bodies.The inclusion bodies were denatured,refoloded and purified by glutathione sepharose 4B affinity chromatography.The purified protein could reacted to positive sera against VSV,indicating that recombinant protein had good antigenicity.