Secreted expression of porcine interferon-gamma gene in Pichia pastoris

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 Pi...

Full description

Saved in:
Bibliographic Details
Published inShengwu gongcheng xuebao Vol. 21; no. 5; p. 731
Main Authors Huang, Zhi-Qing, Hu, Hong-Yu, Chen, Xiao-Ling, Ren, Li-Ming, Lin, Ai-Xing, Chen, Yong-Fu
Format Journal Article
LanguageChinese
Published China 01.09.2005
Subjects
Animals
Electroporation
Interferon-gamma - biosynthesis
Interferon-gamma - genetics
Pichia - genetics
Pichia - metabolism
Recombinant Proteins - biosynthesis
Recombinant Proteins - genetics
Swine - genetics
Online AccessGet more information

Cover

More Information
Summary: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.
ISSN:1000-3061