乙型肝炎病毒HBX基因的克隆、表达和蛋白的纯化  

Expression,Purification and Identification of the Hepatitis B Virus X Protein

作　　者：伍静[1] 鲍向红[1] 杨静谊[1] 罗小楠[1] 张良成[1] 贺超[1] 张毅[1] 吴凯[1] 李维国[1] 

机构地区：[1]第四军医大学训练部,西安710032

出　　处：《科学技术与工程》2012年第14期3319-3322,共4页Science Technology and Engineering

摘　　要：克隆乙肝病毒X基因,并在大肠杆菌中进行表达和纯化。用PCR方法从结乙肝病毒基因组扩增出HBX基因片段,克隆至pMD18-T载体中。序列测定正确后,将其亚克隆到表达载体pProExHTa并在大肠杆菌BL21中表达。表达蛋白经SDS-PAG及Western-blot分析后,亲和层析法纯化蛋白。结果成功克隆了HBX基因,并对其在E.coli中进行了表达。SDS-PAGE及Western blot分析表明表达产物正确。通过IMAC纯化系统获得17 kD纯化蛋白,与文献报道相符。结果成功获得了纯化的HBX蛋白,为进一步研究HBX蛋白与宿主蛋白之间的相互作用奠定基础。To clone the gene of HBX of hepatitis B virus,then express the protein of HBX in Escherichia,Coli BL21（E.coli） and purify the expressed protein,the HBX gene was amplified from the genome of HBV by PCR and cloned into plasmid pMD18-T.The fragment sequenced correctly was subcloned into the expression vector pProExHTa and expressed in E.coli BL21.The expressed protein was identified by SDS-PAGE analysis and Western blot method,and subsequently purified it by His-tag purification system.It is resulted that gene of HBX was successfully cloned and expressed in E.coli,and the expressed protein was identified correctly by SDS-PAGE analysis and Western blot analysis.It is conclused that HBX was successfully expressed and purified,for the further study HBX protein and protein host of interaction between the lay the foundation.

关 键 词：乙肝病毒 HBX 表达 纯化 

分 类 号：R373.21[医药卫生—病原生物学]