Expression And Immunological Analysis Of Yanbian Strain VP3 Gene From Gosling Plague Virus

Gosling Plague is caused by the gosling plague virus. It is an acute or subacute, high degree of contact and septic infectious diseases which major infracts 4-20 days goslings and muscovy ducklings. The main characteristics are acute enteritis and the inflammation of parenchymal organs such as liver, kidney, heart and so on. Short course, spread fast and high mortality of the disease. It is one of the most important infectious diseases which harm the healthy development of goose industry.Gosling plague virus is a member of Parvoviridae, Parvovirus. It is an self-replicating parvovirus. Virus genome is single stranded, linear DNA, the length nucleotide sequence is 5. 1kb. It contains two open reading frames (ORF), the left ORF (LORF) code for non-structural proteins NS1 and NS2; right ORF (RORF) code for three kinds of structural proteins (VP), the VP1, VP2, VP3. VP3 is the major capsid protein and an important protective antigen of the virus, and can induce antibodies with neutralization. Therefore, gosling plague virus VP3 gene is very important in the diagnosis and prevention of Gosling Plague.According to the published gosling plague virus B strain genome sequence in GenBank, we designed a pair of specific primers for the conservative region of VP3 gene. We did PCR amplification of gosling plague virus genomic DNA isolated from Yanbian strain, we got 1457bp gosling plague VP3 gene fragment, and compared it with the published gosling plague virus B strain. The results showed that homology is 96.8%. People proved that the gene fragment is gosling plague VP3 gene. The VP3 gene was cloned into the prokaryotic expression vector pGEX-4T-1. After PCR and restriction endonuclease, it proved that we successfully constructed the expression vector pGEX-VP3. We used 1 mmol/L of IPTG induced the constructed expression vector, and expressed that in E. coli BL21. The expression peak in 8 h. The molecular weight of fusion protein pGEX-VP3 is 81 ku, the majority of proteins are inclusion bodies form. After analysis of Western-Blotting, the fusion protein and gosling plague positive serum had specific reaction, the fusion protein had a better reactogenicity.This study laid the foundation for further study on the molecular characteristics of gosling plague virus. It prepared for the preliminary work on the diagnostic reagents and genetic engineering vaccine of gosling plague.