hper1基因3′端UTR区双荧光素酶报告系统构建  

Construction of dual-luciferase reporter assaysystem of hper1 gene 3′UTR

作　　者：赵溪岩[1,2,3] 成姝婷[1,2,3] 勾洵[1,2,3] 王正荣[1,2,3] 肖静[1,2,3] 郭慧玲[1,2,3] 汪宇辉[1,2,3] 

机构地区：[1]四川大学华西基础医学与法医学院生物医学工程研究室卫生部时间生物学重点实验室,四川成都610041 [2]四川大学生物治疗国家重点实验室,四川成都610041 [3]四川大学"985工程--创新药物研究科技创新平台",四川成都610041

出　　处：《四川生理科学杂志》2013年第1期4-6,共3页Sichuan Journal of Physiological Sciences

基　　金：国家自然科学基金(编号:41074131)

摘　　要：目的:构建hper1基因3′端UTR区双荧光素酶报告系统pmiR-RB-REPORTTM-hper1-3′UTR(PMIR-3′UTR),检测其表达,为研究hper1基因的表达调控及microRNA调控靶基因的结合位点提供基础平台。方法:用PCR扩增的方法提取hper1基因3′UTR序列,并连接至pmiR-RB-REPORTTM(PMIR)双荧光素酶报告载体的多克隆位点,测序验证插入序列并转染入A549细胞检测其荧光素酶活性。结果:测序结果表明PMIR-3′UTR的插入序列正确,在转染入A549细胞后其荧光素酶能正常表达。结论:PMIR-3′UTR双荧光素酶报告系统构建成功。Objective： To construct pmiR-RB-REPORT^TM- hper1-3′UTR（PMIR-3′UTR）of hperl and observe the expression of it in human lung cancer cell line A549. To provide effective information of following expression regulation of hperl and regulation of target genes binding sites by rnieroRNA. Methods： Amplification of the fragment containing 3′ UTR of hperl gene by PCR and inserted it into pmiR-RB-REPORT^TM （PMIR） vector. The activity was assayed by measuring luciferase after PMIR-3′UTR was transfected into A549 cells. Results： The sequence of hperl 3′UTR was confirmed by sequencing, and the dual luciferases could normally express when PMIR-3′UTR was transfected into A549 ceils. Conclusion： PMIR-3′UTR was successfully established.

关 键 词：hper1基因 3′UTR区域 PMIR-3′UTR 

分 类 号：R346[医药卫生—基础医学]