The Renaturation and Activity Study of LexA From Pseudomonas aeruginosa / 中国生物工程杂志

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 G25 gel column.The renaturation result were detected by the native polyacrylamide gel electrophoresis and RPHPLC.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 RPHPLC.Gel retardation experiments shows that the renatured LexA have satisfactory biologic activity.