mtHSP70/HSV-TK双基因真核共表达质粒的构建 Construction of Eukaryotic Coexpression Vector Containing Mycobacterium Tuberculosis Heat Shock Protein 70 and Herpes Simplex Virus Thymidine Kinase Genes 【作者】 曾曙光; 刘启才; 王素文; 徐平平; 章锦才; 张积仁;
【Author】 ZENG Shu-guang1,LIU Qi-cai,WANG Su-wen,et al.1Guangdong Provincial Stomatological Hospital & the Affiliated Stomatological Hospital of Southern Medical University,Guangzhou 510280,China
【机构】 广东省口腔医院; 南方医科大学附属口腔医院口腔颌面外科; 广州医学院实验医学研究中心; 南方医科大学珠江医院肿瘤中心;
【摘要】 目的构建结核杆菌热休克蛋白70(mycobacterium tuberculosis heat-shock proteins 70,mtHSP70)和自杀基因单纯疱疹病毒-胸苷激酶(herpes simplex virus-thymidinekinase,HSV-TK)双基因真核共表达质粒pCMV-mtHSP70-IRES-TK。方法登录Genbank查询HSV-TK和mtHSP70基因mRNA序列,应用引物设计软件DNA club分别设计扩增HSV-TK和mtHSP70基因全长cDNA的特异引物。以pcDNA3-TK质粒或mtHSP70质粒DNA为模板,分别采用各自的引物,用prime STAR HS DNA Polymerase进行聚合酶链反应(polymerase chain reaction,PCR),获得HSV-TK和mtHSP70基因cDNA片断,连接到T载体pMD18-T上,转化大肠杆菌TG1后用HSV-TK和mtHSP70基因的特异引物进行菌落PCR,确定含有阳性mtHSP70或HSV-TK重组质粒并将阳性pMD18T-TK重组质粒和pMD18T-mtHSP70重组质粒... 更多还原
【Abstract】 Objective To construct an eukaryotic plasmid PCMV-mtHSP70-IRES-TK that contains and expresses both mycobacterium tuberculosis heat shock protein 70(mtHSP70) gene and suicide gene HSV-TK for the experiments.Methods Login on genbank and search for mRNA sequences of HSV-TK and mtHSP70 genes.Specific primers for the amplification of full length cDNA of HSV-TK and mtHSP70 were designed by primer designing software DNA club.pcDNA3-TK plasmid and mtHSP70 plasmid DNA were used as template.PCR was perfor... 更多还原
【关键词】 mtHSP70/HSV-TK基因; 真核表达; 免疫治疗; 肿瘤; 【Key words】 mtHSP70/HSV-TK gene; Eukaryotic expression; Immunotherapy; Tumor;
【基金】 广东省社会发展项目(2009B030801264)
【文献出处】 广东牙病防治, Journal of Dental Prevention and Treatment, 编辑部邮箱 , 2011年02期
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