pSUPER介导的RNAi抗乙肝病毒S基因的研究.pdfVIP

生物技术通报 · 研究报告 · BIoTECHNoLoGY BULLETIN 2008年第 1期 pSUPER介导的RNAi抗乙肝病毒S基因的研究 张敬武， 栾慎顺z杨倬。刘长梅。 张景海’ (一沈阳药科大学，沈阳 110016；：沈阳农业大学，沈阳 110161； 中国科学院微生物研究所，北京 100101) 摘 要： 目的 研究RNAi抗乙肝病毒的作用。方法 设计并合成一段针对HBV S基因的干扰序列及一段无关 的序列，克隆进pSUPER栽体中，分别构建pSUPER—HBS、pSUPER—nonseme栽体，然后将pSUPER、pSUPER-HBS、 pSUPER-nonsense转染进HepG2．2．15细胞中，用ELISA方法对细胞上清中HBsAg、HBeAg进行检测。尾静脉注射HBV 转基因小鼠，取血清检测。结果 pSUPER-HBS所表达的siRNA成功的抑制了HepG 2．2．15上清及HBV转基因鼠血清 中的HBsN~、HBeAg，抑制率分别达到70％、51％vX及60％、42％。结论 针对HBV S区的siRNA能明显抑制HBV的复制。 关键词： 乙肝 乙肝病毒 RNA干扰 s抗原 Inhibition of Hepatitis B Virus by pSUPER siRNAs Targeted to Hepatitis B virus S gene Zhang Jingwu Luan Shenshun2 Yang Zhuo。 Liu Changmei。 Zhang Jmghai (1Shenyang Pharmaceutical University，Shenyang llO016；2Shenyang Agrictdtural University，Shenyang 110161； 31nstitute of Microbiolog)，Chinese Acamy of Sciences，Bering 100101) Abstract： objective Study the effect of siRNA which can intibit repfication of Hapatitis B virus．Methods Design and synthesize a sequence which target Hapatitis B virus S gene，and a nonsense sequence，cloned into pSUPER． pSUPER、pSUPER—HBS and pSUPER-nonsense were transefected into HepG 2．2．15．The plasmids were injected into HBV mouse．．ELISA was used to detected the change of HBsAg and HBeAg Results The levels of HBsAg and HBeN~ were dramatically decreased by pSUPER-HBS．HBsAg and HBeAg were reduced by 75％ and 48％ in HepG2．2．15 supematant，60％ and 42％ in HBV mouse’S serum respectively．Conclusion pSUPER-HBS、pSUPER-nonsense can effectively inhibit replication of HBV in vitro． Key words： Type B hepatitis Hapatitis B virus RNA interference S antigen 前言 乙型肝炎是由乙肝病毒(Hapatitis B virus，HBV)引起的、以肝脏炎性病变为主并可引起多器官损害的 一 种传染病…。HBV进入细胞后，L(一)链被转录成