军事医学2018，Vol.42Issue(1)：34-37,4.DOI:10.7644/j.issn.1674-9960.2018.01.009
m6A修饰RNA结合蛋白YTHDF2的克隆、表达及其活性分析
Cloning,expression and biological activity of m6A binding protein YTHDF2
摘要
Abstract
Objective To construct the recombinant plasmid of YTH domain family 2(YTHDF2)and express it in E.coli in order to obtain YTHDF2 fusion protein that was capable of binding m 6A-modified RNA.Methods The coding region of YTHDF2 gene was amplified by RT-PCR.The recombinant plasmid pET-28a-YTHDF2 was constructed and expressed in E.coli.The fusion protein was purified by Ni2+-NTA resin affinity chromatography, while the fusion protein activity was analyzed by Ni2+-NTA magnetic spheres.Results and Conclusion The recombinant YTHDF2 protein was expressed in E.coli BL21(DE3)and purified.YTHDF2 fusion protein was capable of binding RNA with m 6A-modification. The preparation of YTHDF2 fusion protein provides an essential tool to study the biological function of RNA with m6A-modification.关键词
质粒/重组融合蛋白质类/RNA/YTHDF2蛋白/甲基转移酶类/基因表达调控Key words
plasmids/recombinant fusion protein/RNA/YTHDF2/methyltransferases/gene expression regulation分类
生物科学引用本文复制引用
苏晨,史祥,付汉江,郑晓飞..m6A修饰RNA结合蛋白YTHDF2的克隆、表达及其活性分析[J].军事医学,2018,42(1):34-37,4.基金项目
国家自然科学基金资助项目(91540202,31470782,81773038) （91540202,31470782,81773038）
国家重点研发计划资助项目(2016YFC1303604) （2016YFC1303604）
