Production of biologically-active recombinant goldfish gonadotropins in transgenic rainbow trout

The feasibility of using rainbow trout Oncorhynchus mykiss embryos as an expression system for proteins was investigated. For model proteins, we selected two goldfish gonadotropins (GTHs), follicle-stimulating hormone (FSH) and luteinizing hormone (LH). To produce single-chain goldfish FSH (scgfFSH)...

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Bibliographic Details
Published in:Fish physiology and biochemistry 2003-01, Vol.28 (1-4), p.473-474
Main Authors: Morita, T, Yoshizaki, G, Kobayashi, M, Takeuchi, T
Format: Article
Language:English
Subjects:
Aquaculture
Bioassays
Carassius auratus
Embryos
Fish
Freshwater
Oncorhynchus mykiss
Salmon
Trout
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Summary:The feasibility of using rainbow trout Oncorhynchus mykiss embryos as an expression system for proteins was investigated. For model proteins, we selected two goldfish gonadotropins (GTHs), follicle-stimulating hormone (FSH) and luteinizing hormone (LH). To produce single-chain goldfish FSH (scgfFSH) and LH (scgfLH), cDNAs encoding glycoprotein hormone (GP) alpha and FSH beta were fused in tandem, and cDNAs encoding GP alpha and LH beta were fused in tandem. The fused cDNAs were ligated with beta -actin promoter, and microinjected into fertilized rainbow trout eggs. After 4-days incubation, the embryos were subjected to western blotting and in vitro bioassays. The recombinant proteins produced by the embryos were immunoreactive to antisera against goldfish GP alpha , N-glycosylated, and biologically active. We conclude that scgfFSH and scgfLH were successfully produced in transgenic rainbow trout.
ISSN:0920-1742
1573-5168
DOI:10.1023/B:FISH.0000030632.41332.e8