Expression, denaturation, renaturation and purification of THANK protein
- VernacularTitle:重组THANK诱导表达和变性、复性及纯化
- Author:
Dong WU
;
Feng SHEN
;
Yonghua LOU
;
Min PENG
;
Binghua JIAO
;
Mengchao WU
- Publication Type:Journal Article
- Keywords:
THANK;
recombinant proteins;
gene expression regulation
- From:
Academic Journal of Second Military Medical University
2001;0(09):-
- CountryChina
- Language:Chinese
-
Abstract:
Objective: To prepare highly purified THANK protein. Methods: THANK was efficiently expressed in E.coli as inclusion bodies. After bacteria were lysed under ultrasonication, TE buffer,1% TritonX and 2 mol /L urea was used to efficiently extract inclusion bodies. After denaturation with 8 mol/L urea,THANK was partially purified by Sephacryl S-200 chromatography, and then subsequent refolding step was optimized for a maximal recovery of biological active protein. The renatured THANK was purified to homogeneity with Q Sephadex Fast Flow gel filtration and then desalted by Sephadex G-25 chromatography. Results: The purity of biologically active THANK was above 97% in SDS-PAGE densitometric studies. Conclusion: An effective method of denaturation, renaturation and purification of recombinant THANK is established.
