Expression and analysis of recombinant pIL-18 in Pichia pastoris.
- Author:
Shi-Bin SONG
1
;
Zhi-Zhong JING
;
Guo-Hu CHEN
;
Xiao-Xiao WANG
;
Rui-Qian ZONG
Author Information
1. Key Laboratory of Veterinary Porasitology of Gansu Province/State Key Laboratory of Veterinary Etiological Biology/Lanzhou Veterinary Research Institute, Chinese Acadarny of Agricultural Science, Lanzhou 730046, China.
- Publication Type:Journal Article
- MeSH:
Animals;
Electrophoresis, Polyacrylamide Gel;
Electroporation;
Interleukin-18;
biosynthesis;
genetics;
Pichia;
genetics;
metabolism;
Recombinant Fusion Proteins;
analysis;
biosynthesis;
genetics;
Swine
- From:
Chinese Journal of Biotechnology
2007;23(5):818-823
- CountryChina
- Language:Chinese
-
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.
