Cloning and expression of human manganese superoxide dismutase cDNA in Pichia pastoris.
- Author:
Min LING
1
;
Xiang-Jin LAI
;
Ke XIE
Author Information
1. Department of Biochemistry and Molecular Biology, Guangxi Medical University, Nanning 530021, China. lingmin70@163.com
- Publication Type:Journal Article
- MeSH:
Cloning, Molecular;
DNA, Complementary;
biosynthesis;
genetics;
Humans;
Pichia;
genetics;
metabolism;
Recombinant Proteins;
biosynthesis;
genetics;
Superoxide Dismutase;
biosynthesis;
genetics
- From:
Chinese Journal of Biotechnology
2005;21(3):478-481
- CountryChina
- Language:Chinese
-
Abstract:
Human manganese superoxide dismutase (hMn-SOD) cDNA was amplified by RT-PCR from total RNA of human liver cell (L02), and cloned into yeast expression vector pPIC9K containing AOX1 promoter and the alpha-factor signal peptide sequence. The resultant pPIC9K-MnSOD was transformed to P. pastoris GS115, screened for Mut+ carrying multiple copies of hMn-SOD. The positive transformants were fermented in flasks and induced by 0.5% methanol. After 4 days of methanol induction, the expressed hMn-SOD was up to 32% of the total proteins in the supernatant by SDS-PAGE with specific activity of 247.7 u/mg.
