Fusion expression, purification and bioassay of IFN-gamma inducible protein-10 and thioredoxin gene in E. coli / 生物医学工程学杂志
Journal of Biomedical Engineering
;
(6): 535-539, 2005.
Article
in Chinese
| WPRIM
| ID: wpr-354256
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.
Full text:
Available
Index:
WPRIM (Western Pacific)
Main subject:
Thioredoxins
/
Recombinant Proteins
/
T-Lymphocytes
/
Cell Biology
/
Escherichia coli
/
Chemokine CXCL10
/
Genetic Vectors
/
Metabolism
Limits:
Humans
Language:
Chinese
Journal:
Journal of Biomedical Engineering
Year:
2005
Type:
Article
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