Transgenic expression of L-gulono-gamma-lactone oxidase in medaka (Oryzias latipes), a teleost fish that lacks this enzyme necessary for L-ascorbic acid biosynthesis.
Transfer of the gene for L-gulono-gamma-lactone oxidase, the missing enzyme in L-ascorbic acid biosynthesis in scurvy-prone animals, into medaka (Oryzias latipes) was successfully done. The expression plasmid pSVL-GLO, carrying rat liver L-gulono-gamma-lactone oxidase cDNA, was microinjected into the cytoplasm of fertilized eggs during the one-cell stage. Four male F0 fish having the transgene in their germ cells came to maturity, and F1 progeny derived from one of the F0 fish possessed L-gulono-gamma-lactone oxidase activity, indicating that the transgene was functionally expressed in the fish. Genomic Southern blot analysis demonstrated that the transgene existed in both chromosome-integrated and extrachromosomal forms.
Toyohara
H
H
Department of Fisheries, Faculty of Agriculture, Kyoto University, Japan.
Nakata
T
T
Touhata
K
K
Hashimoto
H
H
Kinoshita
M
M
Sakaguchi
M
M
Nishikimi
M
M
Yagi
K
K
Wakamatsu
Y
Y
Ozato
K
K
D000818:Animals; D030801:Animals, Genetically Modified; D001205:Ascorbic Acid / Q000096:biosynthesis*; D001483:Base Sequence; D017931:DNA Primers; D005393:Fish Diseases; D015870:Gene Expression; D050544:L-Gulonolactone Oxidase; D008297:Male; D008969:Molecular Sequence Data; D009990:Oryzias; D016133:Polymerase Chain Reaction; D051381:Rats; D012614:Scurvy / Q000235:genetics / Q000662:veterinary; D013401:Sugar Alcohol Dehydrogenases / Q000096:biosynthesis* / Q000235:genetics*
D017931:DNA Primers; D013401:Sugar Alcohol Dehydrogenases; D050544:L-Gulonolactone Oxidase; D001205:Ascorbic Acid