中国畜牧兽医2017，Vol.44Issue(3)：644-650,7.DOI:10.16431/j.cnki.1671-7236.2017.03.004
KLF3基因3′-UTR区双荧光素酶报告载体及其突变载体的构建与活性鉴定
Construction and Activity Identification of Dual Luciferase Reporter Vector and Mutation Vector of KLF3 Gene 3′-UTR
摘要
Abstract
This study was aimed to construct the dual luciferase reporter vector which contained KLF3 gene 3′-UTR and its mutation vector, and analyze the possible miRNAs which could regulate KLF3 gene expression.In this study,The KLF3 gene 3′-UTR was amplified by polymerase chain reaction (PCR) and inserted in the dual luciferase reporter vector which was digested by Xho Ⅰ and Not Ⅰ;With Targetscan to predict the miRNAs which might interact with KLF3 gene 3′-UTR;293T cells was treated with recombinant vector or its mutation vector or empty vector and miRNA mimics or control by Lipofectamine 2000.The dual luciferase reporter assay system was used to compare the activity of luciferase.The results showed that KLF3 gene 3′-UTR might be the action points of miR-21;Double fluorescent report results showed that miR-21 mimics group (0.6900±0.0144) was decreased 31% than the mutation vector group (1.000±0.0688) and the blank control group (1.000±0.0159) with the luciferase activity of 293T cells.This study successfully constructed KLF3 gene 3′-UTR dual luciferase reporter vector and its mutation vector;Preliminary evidence showed that miR-21 could regulate the expression of KLF3 gene.关键词
KLF3基因/双荧光素酶基因报告载体/miR-21/3′-非编码区Key words
KLF3 gene/dual luciferase reporter vector/miR-21/3′-UTR分类
生物科学引用本文复制引用
翟博,李旭,王春昕,赵云辉,孙铭,张立春,张明新..KLF3基因3′-UTR区双荧光素酶报告载体及其突变载体的构建与活性鉴定[J].中国畜牧兽医,2017,44(3):644-650,7.基金项目
国家863资助项目(2013AA102506) （2013AA102506）
国家绒毛用羊产业技术体系(CARS40-16) （CARS40-16）
国家科技支撑计划子课题(2011BAD28B05-1-5) （2011BAD28B05-1-5）
吉林省博士后科研人员科研项目(0010424) （0010424）
吉林省农业科学院博士后科研项目(2060599) （2060599）
