Cloning,expression and purification of fragile X mental retardation protein
- VernacularTitle:脆性X智力低下蛋白的克隆表达与纯化
- Author:
Jian LIU
;
Ke ZOU
;
Ning ZHU
;
Yan SHEN
- Publication Type:Journal Article
- Keywords:
fragile X mental retardation protein(FMRP);
pET22b(+);
BL21(DE3);
RNA binding
- From:
Basic & Clinical Medicine
2006;0(07):-
- CountryChina
- Language:Chinese
-
Abstract:
Objective Explore the conditions of the cloning,expression and purification of FMRP.Methods The plasmid pET22b(+)-FMR1,constructed by molecular cloning,was transformed into E.coli BL21(DE3) competent cells and induced to express FMRP by IPTG.Recombinant FMRP was purified by affinity chromatography,verified by Western-blot,and tested for its RNA binding ability.Results FMR1 cDNA was successfully cloned into pET22b(+) vector and expressed in E.coli BL21(DE3).A protein with Mr 79 000 was purified and confirmed to be FMRP.This protein retained the RNA binding ability of FMRP.Conclusion We successfully expressed recombinant hFMRP with high purity and activity in E.coli,which provided a reliable material to study the function of FMRP.
