Expression of goat follicle-stimulating hormone analogous gene in Pichia pastoris.
- Author:
Huanli YANG
1
;
Xibiao ZHAO
;
Xiaojuan SONG
;
Jiatong DING
Author Information
1. College of Animal Science and Techiology, Yangzhou University, Yangzhou 225009, China.
- Publication Type:Journal Article
- MeSH:
Animals;
Delayed-Action Preparations;
chemical synthesis;
Electroporation;
Female;
Follicle Stimulating Hormone;
analogs & derivatives;
genetics;
Genetic Vectors;
Goats;
Pichia;
genetics;
metabolism;
Recombinant Proteins;
administration & dosage;
biosynthesis;
genetics
- From:
Chinese Journal of Biotechnology
2008;24(3):409-414
- CountryChina
- Language:Chinese
-
Abstract:
In order to obtain the long-acting FSH preparation, the single strand long-acting analogous gene FSHbeta-CTP-alpha was successfully constructed by the C-terminal peptide(CTP) of carboxyl-terminal region of human chorionic gonadotropin with the goat FSHalpha-subunit and beta-subunit genes, then it was inserted into pPIC9K vector. The recombinant plasmid pPIC9K FSHbeta-CTP-alpha was transformed into Pichia pastoris GS115 by electroporation. The multi-copy inserts His+Mut+ were gained by the screening of phenotype and hyper-resistance to G418. After methanol induction, the supernatant was analysised by SDS-Polyacrylamide Gen Electrophoresis and Western blot. The results show that the transformants of FSHbeta-CTP-alpha could express the objective protein successfully and the molecular weight is about 29 kD. The concentration of supernatant was detected by Radio-immunoassay and the average expression of multi-inserts is 91.849 mIU/mL and the low-inserts is 37.419 mIU/mL. The expression of multi-inserts is higher than the low-inserts significantly. This research lay the foundation for studying the structure of FSH and the production of long-acting FSH preparation.
