江苏大学学报（医学版）2014，Vol.24Issue(2)：122-125,4.DOI:10.13312/j.issn.1671-7783.y130121
Bcl-6基因3'UTR双荧光素酶报告质粒的构建及其与miR-346靶向关系的验证
Construction of Bcl-6 gene 3'UTR dual luciferase reporter vector and targeting verification between Bcl-6 and miR-346
摘要
Abstract
Objective:To construct a luciferase reporter vector containing the 3'untranslated region (3' UTR) of Bcl-6 gene and measure the correlation between Bcl-6 and miR-346.Methods:The miRNA targeting Bcl-6 3'UTR was predicted by miRanda and PicTar.The synthetic 3'UTR fragment of Bcl-6 was cloned into PsiCHECK-2 reporter vector.The luciferase reporter vector and miRNA mimics were transfected into 293T cells.The relative luciferase activity was detected.Results:PicTar and miRanda database shared the results that miR-346 have the complementary binding sites with 3'UTR of Bcl-6.Results of double enzyme digestion and DNA sequencing showed that sequence of luciferase reporter vector was correct.The luciferase activity of reporter vector treating miR-346 was decreased observably about 66％.Conclusion:The luciferase reporter vector containing the 3'UTR of Bcl-6 was constructed successfully.关键词
Bcl-6基因/微小RNA/3'非编码区/荧光素酶报告质粒Key words
Bcl-6 / microRNA / 3 'untranslated region / luciferase reporter vector分类
医药卫生引用本文复制引用
陈娟,袁靖,吴晶晶,田洁,汤新逸,芮棵,田新宇,张悦,刘海冰..Bcl-6基因3'UTR双荧光素酶报告质粒的构建及其与miR-346靶向关系的验证[J].江苏大学学报（医学版）,2014,24(2):122-125,4.基金项目
国家自然科学基金资助项目(81072453,31170849,30972748) （81072453,31170849,30972748）
江苏省自然科学基金资助项目(BK2011472) （BK2011472）
江苏省普通高校研究生科研创新计划项目(CXLX11_0608) （CXLX11_0608）
江苏省卫生厅医学科研基金资助项目(Z201313) （Z201313）
江苏省“青蓝工程”项目(苏教师[2010]27号) （苏教师[2010]27号）
