[Gene cloning and immunogenicity analysis of the structural proteins VP1-VP4 of enterovirus 71].

Objective To clone the genes encoding the structural proteins VP1-VP4 of enterovirus 71 and investigate the immunogenicity of the expressed recombinant proteins.Methods The VP1-VP4 cDNAs were amplified by RT-PCR from the extracted viral RNA and cloned into pMD19-T vector.The cloned VP1-VP4 genes were then inserted into the multi-cloning sites of plasmid pQE30a and expressed in E.coli M15 with IPTG induction.After washing with 8 mol/L urea and purification with Ni-affinity chromatography,the recombinant proteins obtained were tested for immunogenicity by Western blotting and ELISA using rabbit antisera against enterovirus 71 and Coxsackie Virus A16.Results The recombinant VP1-VP4 proteins were highly expressed in E.coli M15 and the purified proteins could be specifically recognized by the rabbit sera against enterovirus 71.Conclusion The expressed enterovirus 71 structural proteins show good immunogenicity and can be used for developing enterovirus 71 vaccine and detection kits.