Cloning,expression and biological activity of m6A binding protein YTHDF2
10.7644/j.issn.1674-9960.2018.01.009
- VernacularTitle:m6A修饰RNA结合蛋白YTHDF2的克隆、表达及其活性分析
- Author:
Chen SU
1
;
Xiang SHI
;
Han-Jiang FU
;
Xiao-Fei ZHENG
Author Information
1. 安徽医科大学研究生院
- Keywords:
plasmids;
recombinant fusion protein;
RNA;
YTHDF2;
methyltransferases;
gene expression regulation
- From:
Military Medical Sciences
2018;42(1):34-37
- CountryChina
- Language:Chinese
-
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.