Cloning, Sequence Analysis And Prokaryotic Expression Of VP3 From Goose Parvovirus

The Goose parvovirus is a member of the parvovirus genus of the parvoviridae. It can cause acute infectious disease characterized by acute enteritis and inflammation of liver, kidney and heart in goslings. It has a high mortality, which mostly threaten the goose industry. Goose plague was first reported by Fang Ding-yi in China in 1956 , followed by several reports of this disease and the virus isolate in many countries. VP3 is the main structural protein, so the sequence analysis and expression of VP3 genes are very important to prevent and cure of this disease.A pair of primer (GF/GR) corresponding to the same region of VPS gene sequence published by Zadori.ect, was used to amplified VP3 gene by the Polymerase Chain Reaction (PCR), The PCR products was cloned into pMD18-T vector, transferred the recombinant plasmid into complete cell TG I , Then isolated the recombinant plasmid. The VP3 gene fragment was identified by restriction endonuclease and PCR, then sequenced and analyzed with DNAman software. The result of sequence analysis showed that VP3 gene from different virus strains share the high homology from95.64 percent to 99.81 percent, and there were only difference of five amino acids between these virus isolate. The VP3 gene has 1605bp and encode 534 amino acid.Another pair of primers are designed by the same way of the first pair, VP3 genes were amplified by PCR, the PCR product was cloned into the plasmid PGEX-6P-1, constructed the prokaryotic express vector PGEX-6P-VP3, the latter was transferred into the competent cell BL21, After induced by IPTG, the BL21 cell succeeded in expressing fusion GST-VP3, SDS-PAGE showed that the molecular weight of the fusion GST-VP3 is 8.8+104ku, VP3 is 5.8 +104ku.This study provided the molecular base of conservation in VP3 gene and single serology of GPV, supported the necessary condition for the further study of the physical and chemical character of VP3. This study also gave the proof of taxonomy and evolution of GPV, lay a foundation for gene vaccine and molecular diagnosis reagent...