医学分子生物学杂志2018，Vol.15Issue(2)：93-95,3.DOI:10.3870/j.issn.1672-8009.2018.02.007
一种基于RNA结合蛋白的RNA loop检测方法
A Novel Method for RNA Loop Detection Based on RNA Binding Pro-teins
摘要
Abstract
Objective To explore a novel method for detection of intracellular RNA loop. Methods The protein-RNA complex was cross-linked within living cells.Primers at the different sites of RNA were designed after RNA immunoprecipitation,and qPCR was used to indirectly detect the binding level of RNA binding proteins.Moreover,the nucleic acid fragments were ultrasonically broken after cross-linking of protein-RNA complexes, and adjacent RNA ends were ligated.Inverse PCR was employed to directly detect the RNA loop.Results The binding level of RNA binding proteins was relatively higher at both 5′-and 3′-ends than at the middle segment of RNA,which in-dicated that there was a possible RNA loop.Inverse PCR confirmed the existence of endogenous RNA loop by revealing the long-range interaction of RNA fragments. Conclusion Detection of RNA loop after cross-linking of protein-RNA complexes in living cells holds promises to be used to examine the advanced structure and function of RNA.关键词
RNAloop/RNA结合蛋白/RNA免疫沉淀/反向PCRKey words
RNA loop/RNA binding protein/RNA immunoprecipitation/inverse PCR分类
军事科技引用本文复制引用
苏中静,张田伟,刘广宇,黄东阳..一种基于RNA结合蛋白的RNA loop检测方法[J].医学分子生物学杂志,2018,15(2):93-95,3.基金项目
广东省创新强校工程人才培育项目计划(No.2016 KTSCX044),汕头市科技计划项目(No.2016-8)This work was supported by grants from the Innovation Enhancing College Project of Guangdong Province(No.2016 KTSCX044),and the Science and Technology Planning Project of Shantou(No.2016-8) （No.2016 KTSCX044）
