Expression and analysis of recombinant pIL-18 in Pichia pastoris / 生物工程学报
Chinese Journal of Biotechnology
;
(12): 818-823, 2007.
Artigo
em Chinês
| WPRIM
| ID: wpr-327941
ABSTRACT
The porcine IL-18 gene was amplified from recombinant plasmid pGEM-IL-18 by PCR, then the pPIC9K-IL-18 of fusion expression vector was constructed by inserting IL-18 fragment,and was transformed to GS115 by electroporation, multi-copy recombinant strains were screened by G418. The expression of recombinant fusion protein was induced by methanol, SDS-PAGE was used to analyze expression product, fusion protein was purified by Sephadex G-100 column, bioactivity of IL-18 was measured by MTT assays. Experiment results show fusion protein of pIL-18 secreted by GS115,expression reaches the secretion peak of 160 mg/L at 72 h. We have expressed and purified successfully the recombinant pIL-18 with obvious biological activity in Pichia pastoris.
Texto completo:
DisponíveL
Índice:
WPRIM (Pacífico Ocidental)
Assunto principal:
Pichia
/
Suínos
/
Proteínas Recombinantes de Fusão
/
Eletroporação
/
Interleucina-18
/
Eletroforese em Gel de Poliacrilamida
/
Genética
/
Metabolismo
Limite:
Animais
Idioma:
Chinês
Revista:
Chinese Journal of Biotechnology
Ano de publicação:
2007
Tipo de documento:
Artigo
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