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载体表达siRNA抑制HBV复制与基因表达
引用本文:吴浩,邬开朗,朱应,吴建国,杨复华.载体表达siRNA抑制HBV复制与基因表达[J].武汉大学学报(理学版),2006,52(4):466-470.
作者姓名:吴浩  邬开朗  朱应  吴建国  杨复华
作者单位:武汉大学,生命科学学院/病毒学国家重点实验室,湖北,武汉,430072
基金项目:国家自然科学基金;教育部高等学校博士学科点专项科研基金
摘    要:用载体表达siRNA的方法并通过RNA干扰达到抑制乙型肝炎病毒(HBV)的复制及其基因的表达.针对HBV的S基因选取4个靶位点,并将目的基因与荧光素酶报道基因构成快速筛选系统,从4个位点中筛选到一个有效的作用位点.用半定量RT-PCR、ELISA、Western blot和荧光定量PCR等方法对筛选到的有效位点进行了验证,结果表明,siRNA成功降解了HBVS基因的mRNA,降低了S蛋白的表达水平,有效地抑制了HBV的复制.

关 键 词:RNA干扰  小分子干扰RNA  乙型肝炎病毒
文章编号:1671-8836(2006)04-0466-05
修稿时间:2006年2月20日

Inhibition of Hepatitis B Virus Replication and Gene Expression through Vector Mediated Small Interfering RNA
WU Hao,WU Kailang,ZHU Ying,WU Jianguo,YANG Fuhua.Inhibition of Hepatitis B Virus Replication and Gene Expression through Vector Mediated Small Interfering RNA[J].JOurnal of Wuhan University:Natural Science Edition,2006,52(4):466-470.
Authors:WU Hao  WU Kailang  ZHU Ying  WU Jianguo  YANG Fuhua
Abstract:In this study,we constructed a fast screening system with target gene and luciferase reporter gene.Four siRNA molecules were designed to target the HBV S gene and investigated the effect of short interfering RNA on HBV viral replication and its gene expression.Results indicated that one out of the four siRNA molecules designed could inhibit the expression of HBs gene successfully based on semi-quantitative RT-PCR,ELISA,western blot and fluorescencequantitative PCR analyses.In addition,the selected siRNA was able to reduce the amount of HBV core associated DNA significantly,an intracellular replicative intermediate.Therefore,it provides a powerful tool for the study of gene function and implicates a potential application in the treatment of viral infection.
Keywords:RNA interference  small interfering RNA  hepatitis B virus
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