原核表达系统T7RNA聚合酶_启动子在真核细胞中表达目的基因的实验的研究.pdfVIP

21 2 Vol.21 No.2 2005 3 ChineseJournal of Biotechnology March 2005 T7 RNA Expression of Target Gene in Eukaryotic Cells Driven by Prokaryotic T7 Promoter and its RNA Polymerase * 袁志刚, 张进平, 储以微, 王 缨, 徐 薇, 熊思东 * YUAN ZhiGang, ZHANG JinPing, CHU YiWei, WANG Ying, XU Wei and XIONG Si ong , 200032 Department of Immunology of ShanghaiMedical College of Fudan University , Key Laboratory of Molecular Medicine of Ministry fo ducation, Centerf or Gene Immu nization and Vaccine Research, Shanghai 200032, China 将目前高表达水平强大的原核表达系统之一T7 RNA 聚合酶启动子表达系统通过 一系列改进引入真核 胞通过 转染真核 胞实验表明,采用真核启动子 CMV 调控T7 RNA 聚合酶的表达和在T7启动子下游插入 EMCV IRES 序列两种解决 方案能使该原核表达系统在真核 胞高效表达目的基因, 且能适应不同的真核 胞环境, 是一良好的 胞类型非依赖的表达 体系 T7 RNA 聚合酶, T7 启动子, 原核表达系统, 真核 胞 R39211 A 2005) Abstract To enhance the efficiency of the expression of target gene in eukaryotic cells, one of the strongest prokaryotic expres sion systems, the T7 RNA polymerase and T7 promoter, was introduced into eukaryotic cells. A duelplasmid gene expression system of T7 bacteriophage componentswas developed; one containing the T7 phage RNA polymerase gene under the control of eukaryotic promoter CMV (pCMVT7pol) andtheother (pT7IRES) containing theT7promoter andT7 terminator aswell asEM CV IRES. To test the feasibility of this plasmid system for eukaryotic expression, hepatitis B virus envelop HBV preS2S was usedto construct pT7IRESHBs. The target geneswere expressed efficiently by the eukaryonizedprokaryotic expression system in a variety of the cells indicating C2Cl2, SP20, NIH3T3 andBALBc 3T3, suggesting the potential applications of the expression system in gene therapy and gene immunization. ey words