丙型肝炎病毒p7蛋白上调去唾液酸糖蛋白受体1基因的表达  

Up-regulation of hepatitis C virus p7 on ASGPR1 gene promoter

作　　者：郭江[1,2] 成军[1] 洪源[1] 王琦[1] 邢卉春[2] 毛羽[1] 

机构地区：[1]首都医科大学北京地坛医院传染病研究所,北京市100015 [2]首都医科大学北京地坛医院内五科

出　　处：《中华实验和临床感染病杂志（电子版）》2010年第2期6-8,共3页Chinese Journal of Experimental and Clinical Infectious Diseases(Electronic Edition)

基　　金：国家自然科学基金(30371288)

摘　　要：目的探讨HCVp7对去唾液酸糖蛋白受体1(ASGPR1)启动子转录的调节作用。方法据生物信息学确定ASGPR1的启动子区域,聚合酶链反应(PCR)扩增ASGPR1p,克隆至真核报告载体pCAT3-Basic中,构建pCAT3-ASG-PR1p报告载体;以该质粒转染肝癌细胞系HepG2,用酶联免疫吸附试验(ELISA)检测氯霉素乙酰转移酶(CAT)的表达活性;并与pcDNA3.1(-)-HCVp7共转染HepG2细胞系,用ELISA检测CAT的表达活性。结果成功获得ASGPR1p启动子的正确克隆。pCAT3-ASGPR1p和pcDNA3.1(-)-HCVp7共转染HepG2细胞的CAT表达活性是pCAT3-Basic空载体的7.7倍,是pCAT3-ASGPR1p的2.4倍。结论 ASGPR1启动子有顺式激活下游基因的活性,HCVp7对ASGPR1基因的表达有上调作用。Objective To investigate the transregulating effect of HCV p7 on ASGPR1 gene promoter.Methods Polymerase chain reaction（PCR）technique was employed to amplify the coding sequence of ASGPR1 promoter from HepG2 genomic DNA and the product was subcloned into pCAT3-Basic by Kpn Ⅰ and Bgl Ⅱ,named pCAT3-ASGPR1p.pCAT3-ASGPR1p was transfected into the hepatoblastoma cell line HepG2 and cotransfected with pcDNA3.1（-）-HCV p7 by FuGENE6 transfection reagents.The HepG2 cells transfected with pCAT3-Basic were used as negative control.The activity of CAT in transfected HepG2 cells was detected by an ELISA kit after 48 hours,which reflected the transregulating function of pcDNA3.1（-）-HCV p7 to ASGPR1 gene promoter.Results The report vector pCAT3-ASGPR1p was constructed and confirmed by restriction enzyme digestion and sequencing.The expression of CAT in HepG2 cells cotransfected with pCAT3-ASGPR1p and pcDNA3.1（-）-HCV p7 were 7.7 times high as that of pCAT3-Basic and 2.4 times high as that of pCAT3-ASGPR1p.Conclusions HCV p7 can transregulate ASGPR1 promoter and can up-regulate the expression of ASGPR1 gene.

关 键 词：丙型肝炎病毒p7蛋白 去唾液酸糖蛋白受体1 基因启动子 

分 类 号：R346[医药卫生—基础医学]