Fish eggs as bioreactors: the production of bioactive luteinizing hormone in transgenic trout embryos.
We demonstrated the production of goldfish luteinizing hormone (gfLH) by the use of 4-day-old rainbow trout embryos as novel bioreactors. This expression system has several advantages: target proteins can be rapidly expressed at low cost, and recombinant proteins can be synthesized at low temperatures and can undergo complex post-translational modifications (PTMs). An expression vector containing gfLH cDNA was microinjected into fertilized trout eggs. After 4 days of incubation at 10 degrees C, transgenic embryos were harvested and glycosylated recombinant gfLH was recovered, which stimulated testosterone production in testicular fragments from the goldfish. This is the first report on the successful production of bioactive recombinant gonadotropin originated from cyprinid. Further, these results demonstrate that trout-embryo bioreactors are a potentially powerful tool for the production of functional recombinant proteins.
Morita
Tetsuro
T
Department of Marine Biosciences, Tokyo University of Marine Science and Technology, Minato-ku, Tokyo 108-8477, Japan.
Yoshizaki
Goro
G
Kobayashi
Makito
M
Watabe
Shugo
S
Takeuchi
Toshio
T
D000818:Animals; D030801:Animals, Genetically Modified; D019149:Bioreactors; D015153:Blotting, Western; D002846:Chromatography, Affinity; D004625:Embryo, Nonmammalian; D006054:Goldfish / Q000235:genetics*; D007763:Lac Operon; D007986:Luteinizing Hormone / Q000096:biosynthesis*; D008297:Male; D008845:Microinjections; D017686:Oncorhynchus mykiss / Q000196:embryology / Q000235:genetics*; D010063:Ovum; D011994:Recombinant Proteins / Q000096:biosynthesis; D013737:Testis / Q000378:metabolism; D013739:Testosterone / Q000096:biosynthesis; D014162:Transfection
D011994:Recombinant Proteins; D013739:Testosterone; D007986:Luteinizing Hormone