载体表达siRNA抑制HBV复制与基因表达  

Inhibition of Hepatitis B Virus Replication and Gene Expression through Vector Mediated Small Interfering RNA

作　　者：吴浩[1] 邬开朗[1] 朱应[1] 吴建国[1] 杨复华[1] 

机构地区：[1]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

出　　处：《武汉大学学报（理学版）》2006年第4期466-470,共5页Journal of Wuhan University:Natural Science Edition

基　　金：国家自然科学基金资助项目(30570070);高校博士点基金(20040486037)资助项目

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HBV S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.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 fluorescence-quantitative 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.

关 键 词：RNA干扰 小分子干扰RNA 乙型肝炎病毒 

分 类 号：Q939.47[生物学—微生物学]