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Updated: Jun 28, 2026
Expression of Recombinant Proteins in the Methylotrophic Yeast Pichia pastoris
Published on: February 25, 2010
[High level secretory expression of recombinant human interleukin 18 in Pichia pastoris]
Li-li Yang1, Feng Wei, Hong Liu
1Department of Immunology, Key Laboratory of Cancer Prevention and Therapy, Tianjin Medical University Cancer Institute and Hospital, Tianjin 300060, China. yanglili2006@yahoo.com.cn
Aim:
To construct eukaryotic expression vector and express human interleukin 18 (hIL-18) in Pichia pastoris.
Methods:
The gene encoding of hIL-18 was amplification by PCR. The recombinant pPICZaC/hIL-18 was transformed into the Pichia pastoris X-33 strain via electroporation. The high level expression was selected and assayed by the methods of PCR, SDS-PAGE and Western blot. The rhIL-18 was purified by the methods of hydrophobic chromatography and anion exchange chromatography. The bioactivity of it was initially assayed.
Results:
The rhIL-18 was secreted into the supernatant and the concentration reached to 202 mg/L. The rhIL-18 was further identified by Western blot with specific antibody binding activity. The purity of the rhIl-18 reached about 95%. And rhIL-18 can synergistically induce PBMC to produce IFN-gamma with IL-2.
Conclusion:
A rhIL-18 is successfully constructed and expressed in Pichia pastoris. And this contributes to further study of its function and activity.
