Toyohara H, Nakata T, Touhata K, Hashimoto H, Kinoshita M, Sakaguchi M, Nishikimi M, Yagi K, Wakamatsu Y, Ozato K
Department of Fisheries, Faculty of Agriculture, Kyoto University, Japan.
Biochem Biophys Res Commun. 1996 Jun 25;223(3):650-3. doi: 10.1006/bbrc.1996.0949.
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.
将L-古洛糖酸-γ-内酯氧化酶基因（易患坏血病动物体内L-抗坏血酸生物合成中缺失的酶）成功导入青鳉（Oryzias latipes）。携带大鼠肝脏L-古洛糖酸-γ-内酯氧化酶cDNA的表达质粒pSVL-GLO在单细胞阶段被显微注射到受精卵的细胞质中。4条生殖细胞中含有转基因的雄性F0鱼成熟，其中一条F0鱼的F1后代具有L-古洛糖酸-γ-内酯氧化酶活性，表明转基因在鱼体内功能性表达。基因组Southern杂交分析表明，转基因以染色体整合和染色体外两种形式存在。
