Yuan Zhi-Gang, Zhang Jin-Ping, Chu Yi-Wei, Wang Ying, Xu Wei, Xiong Si-Dong
Department of Immunology of Shanghai Medical College of Fudan University, Key Laboratory of Molecular Medicine of Ministry of Education, Center for Gene Immunization and Vaccine Research, Shanghai 200032, China.
Sheng Wu Gong Cheng Xue Bao. 2005 Mar;21(2):182-6.
To enhance the efficiency of the expression of target gene in eukaryotic cells, one of the strongest prokaryotic expression systems, the T7 RNA polymerase and T7 promoter, was introduced into eukaryotic cells. A duel-plasmid gene expression system of T7 bacteriophage components was developed; one containing the T7 phage RNA polymerase gene under the control of eukaryotic promoter CMV (pCMV-T7pol) and the other (pT7IRES) containing the T7 promoter and T7 terminator as well as EMCV IRES. To test the feasibility of this plasmid system for eukaryotic expression, hepatitis B virus envelop HBV preS2/S was used to construct pT7IRES-HBs. The target genes were expressed efficiently by the eukaryonized prokaryotic expression system in a variety of the cells indicating C2C12, SP2/0, NIH3T3 and BALB/c 3T3, suggesting the potential applications of the expression system in gene therapy and gene immunization.
为提高目标基因在真核细胞中的表达效率，最强的原核表达系统之一，即T7 RNA聚合酶和T7启动子，被引入到真核细胞中。构建了一种T7噬菌体元件的双质粒基因表达系统；一个质粒含有在真核启动子CMV控制下的T7噬菌体RNA聚合酶基因（pCMV-T7pol），另一个（pT7IRES）含有T7启动子、T7终止子以及EMCV IRES。为测试该质粒系统用于真核表达的可行性，利用乙型肝炎病毒包膜蛋白HBV preS2/S构建了pT7IRES-HBs。该原核表达系统在多种真核细胞（包括C2C12、SP2/0、NIH3T3和BALB/c 3T3）中能够高效表达目标基因，提示该表达系统在基因治疗和基因免疫方面具有潜在应用价值。
