江苏大学学报（医学版）2016，Vol.26Issue(3)：204-207,4.DOI:10.13312/j.issn.1671-7783.y160052
A KT23′-U TR荧光素酶报告基因载体构建及与miRNA-625靶向关系验证
Construction of the AKT2 3′-UTR luciferase reporter gene vector and verification of the targeted relationship with miRNA-625
摘要
Abstract
Objective:To identify the targeted-regulating relationship between miRNA-625 and AKT2 via constructing luciferase reporter gene vector.Methods:The AKT2 was predicted as miRNA-625 target gene by biological software.The 3′-untranslated regions(3′-UTR)of the wild type AKT2 and mutant AKT2 sequence were cloned into luciferase reporter vector pMIR-Report.The HEK-293T cells were divided into four groups randomly,wild-type plasmid +miRNA-625 group,wild-type plasmid +miRNA negative con-trol group,mutant plasmid +miRNA-625 group,mutant plasmid+miRNA negative control group.The rel-evant plasmids and miRNAs were transfected into groups by Lipofectamine 2000.The luciferase activity was detected by dual luciferase reporter gene system.Results:The recombinant plasmids were identified cor-rectly.Dual-luciferase reporter assay system revealed luciferase activity of wild-type plasmid+miRNA-625 group was significantly lower than that of wild-type plasmid+miRNA negative control group(P<0.05 ). Conclusion:The miRNA-625 inhibited the luciferase activity of AKT2 wild type vector,which indicates AKT2 gene could be targeted regulated by miRNA-625 .关键词
miRNA-625/AKT2基因/荧光素酶报告基因/3′端非翻译区Key words
miRNA-625/AKT2 gene/luciferase reporter gene/3′-UTR分类
医药卫生引用本文复制引用
魏斌,邓霞,卞秀娟,钱粉红..A KT23′-U TR荧光素酶报告基因载体构建及与miRNA-625靶向关系验证[J].江苏大学学报（医学版）,2016,26(3):204-207,4.基金项目
国家自然科学基金资助项目（81370119）；镇江市社会发展项目 （）
