人乳头瘤病毒58型L1壳蛋白在原核细胞中的高效表达及抗体制备

Expression of the human papillomavirus type 58L1 capsid protein in E.coli and preparation of the HPV58 L1 mouse antiserum

目的 人乳头瘤病毒 5 8型 (HPV5 8)是重要的高度致瘤性病毒之一 ,用原核细胞表达HPV5 8L1主要壳蛋白 (McP) ,并制备相应抗体血清 ,为基因工程疫苗研制打下基础。方法 用聚合酶链反应 (PCR)扩增HPV5 8L1完整编码区基因 ,并克隆、测序。构建原核表达载体pRSETB 5 8L1,表达的L1蛋白经SDS PAGE纯化 ,免疫BALB c小鼠。结果 在原核细胞中表达了HPV5 8L1壳蛋白 ,该壳蛋白的相对分子质量为 6 0 0 0 0 ,此蛋白与HPV16L1抗体有交叉反应。获得抗HPV5 8L1壳蛋白特异性抗体 ,抗体滴度为 1∶80 ,并用该抗体鉴定了昆虫细胞中表达的HPV5 8L1壳蛋白。结论 HPV5 8壳蛋白在原核细胞中获得有效表达 ,纯化免疫小鼠后能产生抗HPV5 8L1特异性抗体 ,此抗体可用于鉴定真核细胞表达的HPV5

Objective Human papillomavirus type 58(HPV58)has been shown to be one of the most important highly\|oncogenic,risky HPV types.The present study aimed at investigating the expression of the HPV58 L1 major capsid protein by \%E.coli\% and preparation of HPV58 L1 antiserum. Methods The full length L1 coden region of HPV58 L1 was amplified by PCR,the cloned,sequenced.The expression vector pRSETB58 L1 was constructed to produce HPV58 L1 protein.the protein expressed was purified by SDS\|PAGE,and used to immunize BALB/c mice.Results HPV58 L1 protein was expressed in \%E.coli\%,which has cross reaction with anti\|HPV16 L1 antibody.The mouse anti\|HPV58 L1 antibody was obtained and used to test the HPV58 L1 protein expressed in insect cell.Conclusion HPV58 L1 major capsid protein was efficiently expressed in \%E.coli\%.The mouse anti\|HPV58 L1 specific antibody was prepared,which can be used to test the HPV58 L1protein expressed in eukaryotic cell.