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Production of biologically-active recombinant goldfish gonadotropins in transgenic rainbow trout
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Production of biologically-active recombinant goldfish gonadotropins in transgenic rainbow trout
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Production of biologically-active recombinant goldfish gonadotropins in transgenic rainbow trout
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Journal Article
Production of biologically-active recombinant goldfish gonadotropins in transgenic rainbow trout
2003
Overview
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.
