Modification,Expression and Purification of Human Endotoxin Binding Peptide Gene
- VernacularTitle:内毒素结合肽的改良、原核表达及纯化
- Author:
Ya-Li SUN
;
You-Sheng LIU
;
Hai-Jie YANG
;
- Publication Type:Journal Article
- Keywords:
mEBP Mutation Expression Purification
- From:
China Biotechnology
2006;0(03):-
- CountryChina
- Language:Chinese
-
Abstract:
Objective: To modify EBP(endotoxin binding peptide), clone and express the mutate of EBP gene and gain purified mEBP.Method: mEBPgene was cloned by PCR site-directed mutagenesis. PinpointXa-3/mEBP expression vector was designed to express human mEBP as a fusion protein in BL21 (DE3) pLysS. Digested engineering bacteria by lysozyme and collected inclusion bodies.Fusion protein was purified by Pinpoint TM Xa purification system and cleaved by factorXa,mEBP was purified by RP-HPLC. Results: Mutations at residues 5 and 18(Gln→Lys) was obtained by PCR site-directed mutagenesis, expressed and purified mEBP successfully.Conclusions: Obtaining of purified mEBP lay a foundation for its biological activity research.
