DNA疫苗_____DNA疫苗；乙型肝炎表面抗原；表达

重组乙肝表面抗原真核表达质粒的构建及表达

http://www.100md.com 《广东药学院学报》 2001年第4期

DNA疫苗,,DNA疫苗；乙型肝炎表面抗原；表达,1材料,2方法与结果,3讨论,参考文献

     摘要采用限制性内切酶从重组的真核表达质粒pcDNAS2S中分离出经过测序鉴定的乙肝表面抗原中蛋白(preS2+S)基因片段，将其亚克隆于pVAX1真核表达载体，构建编码乙肝表面抗原中蛋白的卡那霉素抗性真核表达质粒pVAXS2S，酶切鉴定后将重组质粒体外转染COS7细胞，ELISA法检测上清液中的HBsAg呈阳性，说明重组质粒pVAXS2S在体外能够有效表达HBsAg，为进一步的体内免疫反应打下基础。

    关键词 DNA疫苗；乙型肝炎表面抗原；表达

    Construction and expression of recombinant eukaryotic expression vector containing hepatitis B virus surface antigen gene

    WU Hong1，LUO Minxin2，CHEN Guangming3

    (1.Department of Biotechnology,South China University of Technology,Guangzhou,Guangdong 510640；2.Jiang Men Wu Yi Hospital of Traditional Chinese Medicine；3.Liver Diseases Institute of Guangzhou Hospital of Airforce)

    Abstract Using restriction enzymes,the gene fragment encoding HBV antigen middle protein (preS2+S) was isolated from the recombinant eukaryotic expression plasmid pcDNAS2S,then subcloned into eukaryotic expression vector pVAX1. In this way,a recombinant kanamycin resistant eukaryotic expression plasmid (pVAXS2S) containing HBV surface antigen gene was constructed. After identified by restriction enzymes digestion,it was then used to transfect COS7 cells. HBsAg was detected in the supernatant by ELISA,suggesting that the recombinant plasmid could produce HBsAg effectively in vitro. This has provided a basis for further immune response research in vivo. ......

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