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. 2007 Sep;23(5):818-23.

[Expression and analysis of recombinant pIL-18 in Pichia pastoris]

[Article in Chinese]
Shi-Bin Song  1 , Zhi-Zhong Jing, Guo-Hu Chen, Xiao-Xiao Wang, Rui-Qian Zong
Affiliations
  • PMID: 18051858

[Expression and analysis of recombinant pIL-18 in Pichia pastoris]

[Article in Chinese]
Shi-Bin Song et al. Sheng Wu Gong Cheng Xue Bao. 2007 Sep.

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.

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