应用RNAi文库筛选HBV复制相关基因  

Screening of Genes Involved in HBV Replication in RNAi Library

作　　者：曾贵利[1] 沈薇[1] 吴进峰[1] 任红[2] 孙航[2] 贺彩霞[2] 唐开福[2] 

机构地区：[1]重庆医科大学附属第二医院消化内科,重庆400010 [2]重庆医科大学病毒性肝炎研究所,重庆400010

出　　处：《中国生物化学与分子生物学报》2008年第12期1165-1169,共5页Chinese Journal of Biochemistry and Molecular Biology

基　　金：重庆市卫生局医学科学技术研究项目(No.06-2-083)资助~~

摘　　要：深入研究HBV复制机理,筛选参与HBV复制的基因,可能为开发抗乙肝病毒新药提供新的靶点.本文拟建立一种筛选HBV复制相关基因的方法:RNAi文库感染HepG2.2.15细胞后,利用免疫磁珠收集HBsAg表达降低的细胞,提取DNA,PCR扩增siRNA编码序列,将PCR产物克隆入T-easy载体,随机挑选克隆测序,发现DDB1基因可能参与HBV复制.本试验建立了一种筛选HBV复制相关基因的方法,为大规模全基因组筛选参与HBV复制的基因奠定了基础.Understanding the molecular mechanism of Hepatitis B virus （HBV） replication is important for designing new agents and devising strategies to manage and prevent HBV infection. A method to screen genes involved in HBV replication was developed. HepG 2.2.15 cells were infected with RNAi library. Cells expressing less HBsAg were isolated using immunomagnetic beads. The siRNA coding sequences were PCR- amplified using DNA extracted from cells expressing less HBsAg as template, and the PCR products were cloned into T-easy vector. After random sequencing, we found that DDB1 might be involved in HBV replication.

关 键 词：RNAi文库 乙型肝炎病毒(HBV) 筛选 

分 类 号：Q51[生物学—生物化学] Q78