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.
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.