Interleukin 18 High Level Expression in E.coli Purification and Renaturation of the Recombinant Protein.
Sheng Wu Hua Xue Yu Sheng Wu Wu Li Xue Bao (Shanghai)
; 32(4): 397-400, 2000.
Article
em En
| MEDLINE
| ID: mdl-12075431
ABSTRACT
Using the total RNA extracted from mitogen-stimulated human peripheral blood mononuclear cells (PBMC) as template, the cDNA of interleukin 18 was amplified by RT-PCR. The cDNA was subsequently cloned into the expression vector pJW2 and sequenced. The recombinant human IL-18 (rhIL-18) was expressed efficiently in inclusion bodies in E.coli with the yield accounting for 20% total bacteria proteins. The inclusion bodies were washed with 2 mol/L urea and rhIL-18 was further purified using Sephadex G-100 column chromatography in 8 mol/L urea. After purification, the purity of rhIL-18 was greater than 90% as judged by SDS-PAGE. The purified rhIL-18 showed significant and dose-dependent IFN-gamma-inducing activity in human PBMC, in the presence of 0.5 mg/L Con A.
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Coleções:
01-internacional
Base de dados:
MEDLINE
Idioma:
En
Revista:
Sheng Wu Hua Xue Yu Sheng Wu Wu Li Xue Bao (Shanghai)
Ano de publicação:
2000
Tipo de documento:
Article
País de afiliação:
China