HPV16 L1蛋白在大肠杆菌中的表达、纯化及免疫原性研究

EXPRESSION, PURIFICATION, AND IMMUNIZATION OF HPV16 L1 PROTEIN IN E.coli

用IPTG诱导目的工程菌 pQE31 HPV16L1/M 15 (pREP4) ,对表达产物进行SDS PAGE和Westernblot分析 ;用表达的L1蛋白免疫BALB/C小鼠得到抗血清后 ,利用真核源性的VLP粗提物验证小鼠抗血清的特异性。利用IMAC金属亲和层析柱纯化L1蛋白。SDS PAGE结果显示表达产物在约 5 7ku处有蛋白条带 ;Westernblot结果证实此条带可与HPV16L1蛋白的单克隆抗体反应 ;纯化后的L1蛋白也同样保留免疫特异性 ;小鼠抗血清可与HPV16L1VLP(病毒样颗粒 )发生特异性反应 ,证实重组表达的L1蛋白具有免疫原性。本实验表明HPV16L1蛋白在工程菌M15 (pREP4)中高效表达 。

In order to obtain prokaryotic expressed HPV16 L1 protein and develop gene engineering vaccine, an engineering E.coli pQE31 HPV16L1/M15(pREP4) was induced with IPTG and analyzed the induced products by SDS PAGE and Western blot. The Ni 2+ affinity chromatography was used to purify the His 6 L1. BAL B/C mice were immunized with the L1 protein. A Mr 57 ku target band was seen in SDS PAGE that has specific reaction with monoclonal antibody of HPV16 L1. The eukaryotic expressed HPV16 L1 VLP was used to confirm the specific antiserum. The antiserum could react with VLP that demonstrated antigen activity of the His 6 L1 protein. Thus the HPV16 L1 protein, which has antigen activity in E.coli , is successfully expressed.