[Expression of goat follicle-stimulating hormone analogous gene in Pichia pastoris].
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.
Yang
Huanli
H
College of Animal Science and Techiology, Yangzhou University, Yangzhou 225009, China.
Zhao
Xibiao
X
Song
Xiaojuan
X
Ding
Jiatong
J
D000818:Animals; D003692:Delayed-Action Preparations / Q000138:chemical synthesis; D018274:Electroporation; D005260:Female; D005640:Follicle Stimulating Hormone / Q000031:analogs & derivatives* / Q000235:genetics; D005822:Genetic Vectors; D006041:Goats; D010843:Pichia / Q000235:genetics / Q000378:metabolism*; D011994:Recombinant Proteins / Q000008:administration & dosage / Q000096:biosynthesis* / Q000235:genetics
D003692:Delayed-Action Preparations; D011994:Recombinant Proteins; D005640:Follicle Stimulating Hormone