基因重组人巨细胞病毒pp150蛋白片段的原核表达及鉴定  

Construction of human cytomegalovirus pp150 gene prokaryotic expression clones and its mechanism

作　　者：郭大东[1] 高雪芹[1] 韩金祥[1] 刘毅[1] 张华宁[1] 

机构地区：[1]山东省医药生物技术研究中心卫生部生物技术药物重点实验室山东省现代医用药物与技术重点实验室,山东济南250062

出　　处：《山东医药》2006年第17期13-14,共2页Shandong Medical Journal

基　　金：山东省科技厅科技攻关计划资助项目(2004GG2202150)

摘　　要：目的以人巨细胞病毒(HCM V)AD 169病毒株基因为模板,经PCR扩增了编码pp150蛋白片段的UL 32基因,转入pM D 18-T克隆载体后经酶切与表达载体pET-11a连接,转入大肠杆菌BL 21,重组大肠杆菌经诱导表达pp150蛋白片段。经SDS-PAGE分析,其相对分子量约为21.5 kD,表达量约占菌体蛋白的16.38%,W est-ern b lot鉴定为阳性。表明成功构建了pp150蛋白片段的表达载体,并成功诱导表达了pp150蛋白,为进一步纯化该蛋白奠定了基础。[Objective] To construct human cytomegalovirus pp150 gene prokaryotic expression clones and produce recombinant antigen for serodiagnosis of HCMV infection. [Methods] One fragment encoding ppl50 antigen epitopes （495-691） were amplified by PCR and inserted into prokaryotic expression vector pET-11a, transferred into the host bacteria BL21 and induced to express pp150 protein, then identified by western blot. [Results] Expression vector pp150-pET-11a clones were expressed effectively and its relative molecular weight was 21.5kD analysed by SDS-PAGE, and its product was 16.38 % in the total protein, the expressed protein was analysed by Western blot. [Conclusions] The pp150 protein is expressed effectively in the BL21strains and lays foundation for further application.

关 键 词：人巨细胞病毒 原核表达 

分 类 号：R78[医药卫生—口腔医学]