Abstract:
Hepatopancreas parvovirus(HPV) is one of the important pathogens of shrimp. The purpose of this study was to express full-length capsid protein(CP) of HPV in Escherichia coli and prepare polyclonal antibodies against CP protein. In this study, the full-length capsid protein gene was synthesized based on the sequence of HPV virus(Korean strain) from Penaeus chinensis, which was subcloned into pSUMO vector and transformed into Rossetta(DE3) strain to induce expression. After obtaining the purified SUMO-CP fusion protein, polyclonal antibodies serum were obtained by immunizing rabbits. The antigen affinity purification chromatography column was prepared by coupling SUMO protein with agarose medium to remove non-specific antibody components, and the serum titer and specificity were further determined by ELISA and Western Blotting. The results indicated that the recombinant plasmid of pSUMO-sHPVCP was successfully constructed. The fusion protein of SUMO-CP was proved successfully expressed and existed in the form of inclusion body in E. coli by SDS-PAGE and WB detection. The purity of CP protein was more than 90% after purification with nickel column. Ten milligrams of SUMO-CP fusion protein was obtained for the preparation of polyclonal antibodies. After removal of cross-reactive antibodies against SUMO by SUMO protein affinity chromatography column, the specificity of anti-serum was enhanced. The yield of polyclonal antibodies was more than 5.4 mg. The titer of anti-CP protein in serum was more than 512 000. WB test showed that the specificity against CP of serum was good. This study laid a foundation for further research on the pathog-enesis, immune agents and diagnostic reagents of shrimp HPV.