Expression Of The VP2 Gene Fragment Of Goose Parvovirus In Prokaryotic System And Preparation Of Its Antiserum

Goose parvovirus (GPV) is the causative agent of Goose plague, which is also named as Derzsy'sdisease. It is one of highly fatal diseases of goslings and Muscovy ducklings aged 4 to 20 days. The diseaseis characterized by acute enteritis and high mortality which is up to 70%—100%. At present, there are norapid diagnosis methods and effective treatment available. The prevention methods of the disease dependmainly on traditional vaccination and diagnosis based on serological and Virology methods, but they are alltime consuming and laborious. A rapid and special diagnosis method and new genetic engineering vaccineare required to detect and control the infection.VP2 is one of the viral structural proteins on the surface of GPV, which can neutralize antigen andproduce antibody against GPV. In earlier study, a field strain of GPV named HG5/82 was isolated fromgoose embryos and sequenced. The result showed that the nucleotide identity between HG5/82 and GPVstrain B was 93.6%.In this study, specific primers for HG5/82 strain were designed to amplify the VP2 gene. Byrecombinant DNA techniques, a 969bp fragment at the 5'-end of VP2 gene of HG5/82 strain wassubcloned into Nco I site of prokaryotic expression vector pPROEXTMHTb. The recombinant plasmid wastransformed into E.coli DH5αand the bacteria were induced with IPTG. SDS-PAGE analysis showed aninduced product band about 36kDa, which was corresponding to the size of the fragment. The recombinantprotein was expressed efficiently in form of inclusion body. The amount of the recombinant protein wasevaluated by densitometric scanning. It indicated that the product was 21.4% of total bacterial protein ofDH5α.The induced bacterial was solubilized by 6mol/L Guanidine hydrochloric acid and purified byProBondTM Resin. The antiserum against the recombinant protein was obtained by injecting the rabbit withfusion protein. Then the antiserum was tested by western blot. As a result, the antiserum had a positivereaction with HG5/82 strain.This study established a foundation and prepared experimental material for the future research about thebioactivity of GPV. It also provided a basis for developing GPV molecular diagnosis reagent and geneticengineering vaccine.