The Renaturation and Activity Study of LexA From Pseudomonas aeruginosa
- VernacularTitle:铜绿假单胞菌SOS反应阻遏蛋白LexA的复性及活性研究
- Author:
Xuan CHEN
;
Shaohui TANG
;
QingBing ZHA
;
Hui TANG
;
Fang LIU
;
- Publication Type:Journal Article
- Keywords:
Pseudomonas aeruginosa LexA Renaturation
- From:
China Biotechnology
2006;0(09):-
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To optimize the renaturation procedure of denatured LexA,prepare the repressor LexA from Pseudomonas aeruginosa(PA),which have the satisfactory biologic activity.Methods:The LexA was renatured by the GSH/GSSG dilution method,and the renatured protein were purified by Ni2+ chelate affinity chromatography and gel filtration chromatography,following desalination by Sephadex G25 gel column.The renaturation result were detected by the native polyacrylamide gel electrophoresis and RPHPLC.The immunological activity of all LexA proteins,including the denatured,renatured protein and the renatured protein that was treated with the DTT,were determined by Western blot.Results:The renatured LexA appears both monomer and multimer,which is confirmed by the native polyacrylamide gel electrophoresis analysis and RPHPLC.Gel retardation experiments shows that the renatured LexA have satisfactory biologic activity.
