插入CpG序列的乙肝病毒核心抗原基因真核表达质粒的构建及鉴定  

Construction and identification of eukaryotic expression vector containing hepatitis B virus C antigen gene and CpG motif

作　　者：周健[1] 田德英[1] 章述军[1] 许东[1] 

机构地区：[1]华中科技大学同济医学院附属同济医院感染科,湖北武汉430030

出　　处：《中国现代医学杂志》2008年第20期2962-2964,2968,共4页China Journal of Modern Medicine

基　　金：湖北省自然科学基金资助项目(No:2006ABA139)

摘　　要：目的构建pZeoSV2(+)/CpG-HBcAg(ISS)基因真核表达载体,为研制乙肝治疗性疫苗奠定基础。方法根据乙肝病毒核心抗原基因序列,设计合成3对引物,在引物中引入分别针对人、鼠敏感的CpG片段和不含CpG的片段,用PCR方法从慢性乙肝患者血清DNA中扩增出乙肝核心抗原基因片段,将扩增的产物与真核表达质粒pZeoSV2(+)连接,构建重组体pZeoSV2(+)/CpG-HBcAg(ISS),进行酶切、PCR及测序鉴定。结果乙肝核心抗原基因体外扩增产物大小约为530bp。所构建的pZeoSV2(+)/CpG-HBcAg(ISS)重组体经双酶切及PCR鉴定,与预期片段的大小相符;测序结果与GenBank中收录的HBcAg全长序列一致,表明pZeoSV2(+)/CpG-HBcAg(ISS)真核表达载体构建正确。结论成功地构建了真核重组表达载体pZeoSV2(+)/CpG-HBcAg(ISS),为CpG的功能研究和乙肝治疗性疫苗的研制提供了物质基础。Objective To construct the recombinant eukaryotic expression vector pZeoSV2(+)/ CpG-HbcAg(ISS), which provided the basic material for the development of Hepatitis B virus remedial DNA vaccine.Methods Three couples of primers were designed for PCR according to the known sequence of HBcAg.The CpG fragments sensitive to human or murine and Non-CpG fragments are pulled into primers.The HBcAg gene was amplified by PCR from genome of serumal DNA of chronic HBV patients, and PCR product was subcloned into eukaryot...

关 键 词：乙肝病毒核心抗原 CPG基序 TOLL受体 疫苗 

分 类 号：R392[医药卫生—免疫学]