Secreted expression of porcine interferon-gamma gene in Pichia pastoris.
- Author:
Zhi-Qing HUANG
1
;
Hong-Yu HU
;
Xiao-Ling CHEN
;
Li-Ming REN
;
Ai-Xing LIN
;
Yong-Fu CHEN
Author Information
1. National Laboratory for Agrobiotechnology, College of Biology Science, China Agricultural University, Beijing 100094, China.
- Publication Type:Journal Article
- MeSH:
Animals;
Electroporation;
Interferon-gamma;
biosynthesis;
genetics;
Pichia;
genetics;
metabolism;
Recombinant Proteins;
biosynthesis;
genetics;
Swine;
genetics
- From:
Chinese Journal of Biotechnology
2005;21(5):731-736
- CountryChina
- Language:Chinese
-
Abstract:
The porcine interferon-gamma (PoIFN-gamma) gene, in which the sequence encoding signal peptide was replaced by that of the alpha-factor of Saccharomyces cerevisiae, was cloned into Pichia pastoris expression vector pPIC9K. The recombinant plasmid pPIC9K-alpha-PoIFN-gamma was then transformed into Pichia pastoris GS115 cells by electroporation and stable multicopy recombinant Pichia pastoris strains were selected by G418 resistance. Two recombinants of multiple inserts were obtained. SDS-PAGE and Western blot assays of culture broth from a methanol-induced expression strain demonstrated that recombinant PoIFN-gamma, 17kD and 23kD proteins, were secreted into the culture medium. Target proteins, 60% of total proteins, were obtained in the culture medium at the concentration of 108 mg/L. This is the first secreted expression of porcine interferon-gamma gene in Pichia pastoris.
