Expression of human oncostatin M in the methylotrophic yeast Pichia pastoris and its activity identification
- VernacularTitle:抑瘤素M在毕赤酵母中的表达及活性鉴定
- Author:
Ping, LU
;
Xing, LU
;
Rui-yun, XING
- Publication Type:Journal Article
- From:Bulletin of The Academy of Military Medical Sciences
2001;25(1):39-41,74
- CountryChina
- Language:Chinese
-
Abstract:
Objective: To establish the yeast expression system for oncostatin M (OSM). Methods:The cDNA encoding human OSM was inserted into pPIC9K vector and transformed into Pichia pastoris. The correct phenotype of recombinants was screened through MD and MM auxotrophic culture medium. The OSM mRNA transcription in the P.pastoris and molecular weight were identified with Northern blot and Western blot. MTT method was used to test the activity of OSM in inhibiting the growth of human melanoma cells. Results: Established P.pastoris system could accomplish human OSM secreting expression. The recombinant OSM with a 30 000 molecular weight amounted to 70% of the total protein in medium supernatant. The OSM showed an inhibitory effect on the growth of A375 cells in vitro. Conclusions：The P.pastoris system of OSM was proved effective with a high expressing level. The secreted recombinant OSM present in supernatant of culture medium would be easy to be purified. It may lay the foundation for the large-scale preparation of OSM in future.
