Construction and proeukaryotic expression of a novel VP3 protein of JC virus

Objective To construct proeukaryotic expressive vector of VP3 gene of JC virus,and observe the expression of recombinant protein.Methods The DNA fragment of vp3 was amplified by polymerase chain reaction(PCR),and cloned into pGEM-T vector.After sequencing,the correct DNA fragment was inserted into inducible proeukaryotic expressive vector pET-32a(+) and transformed into E.coli BL21.The protein was induced with IPTG and analyzed with sodium dodecylsulfate-polyacrylamide gel electrophoresis and Western blot hybridization.Results The DNA fragment of vp3 was amplified by PCR.The expressive vector was constructed successfully.After induction with IPTG,recombinant target protein with Mr 59 ku was expressed.Western blot analysis showed that the protein had good antigenicity.Conclusion The recombinant vp3 gene was expressed successfully.These results lay the foundation for studying the immunogenicity and bionomics of VP3 protein.