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- Title
Expression of gonadotropin genes in Manchurian trout Brachymystax lenok and production of recombinant gonadotropins.
- Authors
Eunju Choi; Hyeyeon Ko; Jihye Shin; Mi-Ae Kim; Young Chang Sohn
- Abstract
Manchurian trout Brachymystax lenok is an endangered fish species in East Asia including the Korean peninsula. To establish a method for artificial propagation and to improve our understanding of reproduction in the species, cloning of the pituitary gonadotropin (GTH) subunit cDNAs encoding the Manchurian trout gonadotropin common α (mtGTHα), follicle-stimulating hormone β (mtFSHβ), and luteinizing hormone β (mtLHβ) by polymerase chain reaction, were attempted. The open reading frames of the mtGTHα, mtFSHβ, and mtLHβ encoded 114, 137, and 142 amino acid residues, respectively. The mature peptides showed strikingly high sequence identities to those of salmonid species GTHα2, FSHβ, and LHβ subunits (92–100%). In an examination of tissue distribution of the mtGTHα, mtFSHβ, and mtLHβ, it was revealed that all of the three subunit genes are specifically expressed in the pituitary by reverse transcription–polymerase chain reaction analysis. Next, single-chain recombinant FSH and LH constructs using the mtGTHα, mtFSHβ, and mtLHβ cDNAs were produced. In a transient transfection of the FSH and LH constructs into Chinese hamster ovary (CHO-K1) cells followed by Western blot analysis, the tethered FSH in cell lysates or LH secreted into a medium was detected from 48 to 72 h after transfection. The present study provides a possibility of bioactive GTH production for studies on the reproductive physiology of GTH in the Manchurian trout.
- Subjects
FISH genetics; GENE expression; TROUT; GONADOTROPIN; FISHES; RARE fishes; FISH physiology; FISHERY sciences; FUNCTIONAL genomics
- Publication
Fisheries Science, 2005, Vol 71, Issue 6, p1193
- ISSN
0919-9268
- Publication type
Article
- DOI
10.1111/j.1444-2906.2005.01083.x