Prokaryotic expression and analysis of Scophthalmus maximus reovirus (SMReV) VP5 protein

Scophthalmus maximus reovirus (SMReV) belongs to the Aquareovirus-A in the family Reoviridae. The virus particles are icosahedral in symmetry and have a double-layered capsid which is composed of VP5 protein and VP7 protein. The full-length (2 057 bp) of vp5 was cloned from SMReV genome. The prokaryotic expression plasmid pET32a-vp5 was constructed and transformed into E.coli BL21 (DE3) in order to obtain recombinant protein. After being induced, the fusion protein was expressed as inclusion body with molecular weight of approximately 88 kD. The protein was purified and used to generate anti-SMReV VP5 sera in mice. Western blot analysis shows immune reaction of SMReV VP5 with sera, suggesting that the sera were successfully produced and recognized SMReV VP5 with molecular weight of about 69 kD. Indirect immunofluorescence assay (IFA) suggests that VP5 aggregated as granular structures in the cytoplasm of infected CIK cells.