应用抑制性消减杂交技术筛选HCV p7TP2反式调节基因  

Screening of the target genes transactivated by HCV p7TP2 with suppression subtractive hybridization technique

作　　者：袁菊[1] 成军[1] 洪源[1] 陶明亮[1] 靳亚平[2] 李越[1] 李康[1] 杨延平[2] 

机构地区：[1]北京地坛医院传染病研究所,北京市100011 [2]西北农林科技大学动物科技学院

出　　处：《中华实验和临床感染病杂志（电子版）》2008年第3期153-161,共9页Chinese Journal of Experimental and Clinical Infectious Diseases(Electronic Edition)

基　　金：国家自然科学基金项目(C03011402;C30070689);军队九五科技攻关项目(98D063);军队回国留学人员启动基金项目(98H038);军队十五科技攻关青年基金项目(01Q138);军队十五科技攻关项目(01MB135)

摘　　要：目的应用抑制性消减杂交技术筛选HCV p7TP2反式调节基因。方法采用实时定量PCR验证HCV p7下调p7TP2基因的表达,并应用抑制性消减杂交技术筛选并构建p7TP2下调表达基因的消减文库。用活细胞发光法检测p7TP2基因抑制Huh-7细胞系的增殖。结果经同源性分析p7TP2下调表达的基因主要位于线粒体,并与细胞凋亡有关。p7TP2基因转染的Huh-7细胞的活性被显著抑制。结论p7TP2基因功能的研究为HCV致病机制的研究奠定了基础。Objective To screen the target genes transactivated by HCV p7TP2 with suppression subtractive hybridization technique.Methods Realtime PCR was used to check whether HCV p7 down-regulated expression of p7TP2 and suppression subtractive hybridization technique was used to screen and set up the subtractive library down-regulated by HCV p7TP2.Cell viability was detected by celltiter-Glo luminescent to check whether p7TP2 gene could inhibit generation of huh-7 cells.Results Homological analysis showed p7TP2 down-regulated genes mainly located in the mitochondrion and involved in cell apoptosis.The growth of huh-7 cells transfected with p7TP2 gene was dramatically inhibited compared with the parent huh-7 cells.Conclusions This finding brought some new clues for further study on biological functions of p7TP2 and established a foundation for studying the pathogenesy of HCV.

关 键 词：实时定量PCR 下调表达 细胞凋亡 丙型肝炎病毒 

分 类 号：R346[医药卫生—基础医学]