Fusion expression, purification and bioassay of IFN-gamma inducible protein-10 and thioredoxin gene in E. coli.
- Author:
Gang LI
1
;
Ling TIAN
;
Yuquan WEI
;
Yanjun WEN
;
Fei XIAO
;
Bing YAO
;
Ling ZHANG
;
Ru ZHANG
;
Kai MEI
Author Information
1. Key Laboratory of Biotherapy of Human Diseases of Ministry of Education and Cancer Center, West China Hospital, Sichuan University, Chengdu 610041, China.
- Publication Type:Journal Article
- MeSH:
Chemokine CXCL10;
biosynthesis;
Escherichia coli;
metabolism;
Genetic Vectors;
Humans;
Recombinant Proteins;
biosynthesis;
T-Lymphocytes;
cytology;
Thioredoxins;
biosynthesis
- From:
Journal of Biomedical Engineering
2005;22(3):535-539
- CountryChina
- Language:Chinese
-
Abstract:
Interferon gamma-inducible protein 10, a member of the family of CXC chemokines, is secreted by interferon gamma-stimulated, monocytes, endothelial cells and keratinocytes. Interferon gamma-inducible protein 10 plays an important role in recruiting activated T cells into sites of tissue inflammation. In this experiment, PCR products of Interferon gamma-inducible protein 10 were cloned into prokaryote expression vector pET 32(a) to generate recombinant pET-IP10 with S-Tag at the N-terminus, and expressed successfully in E. coli BL21 (DE3). The total expressed products amounted to 25.3% in all bacterion proteins. pET-IP10 mainly formed inclusion body in E. coli. Soluble recombinant protein accounted for 20% among IP-10 fusion protein. The soluble recombinant proteins were purified by using S-Tag affinity chromatography effectively with purity of over 90%. The chemotaxis biological activity of purified Interferon gamma-inducible protein 10 could specifically exhibit the directional migration of stimulated T cells at concentration of 100 ng/ml. The results indicated that the strategy we used in this experiment was effective for recombinant Interferon gamma-inducible protein 10 production with biological activity.
