GENE-TRANSFER IN TILAPIA (OREOCHROMIS-NILOTICUS)
GENE-TRANSFER IN TILAPIA (OREOCHROMIS-NILOTICUS)
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DOI：
10.1016/0044-8486(88)90354-7
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发表时间：
1988-02-15
期刊：
影响因子：
4.5
通讯作者：
WINNACKER, EL
中科院分区：
文献类型：
--
作者：
BREM, G;BRENIG, B;WINNACKER, EL
本文报道了将人生长激素基因显微注射到罗非鱼胚盘中，并培育出90天活的转基因罗非鱼。使用了产卵后10分钟、21-24小时或40-43小时处理的三种不同的产卵。对于显微注射，使用含有与编码人生长激素（hGH）的结构基因融合的小鼠金属硫蛋白-I启动子（mMT-I）的线性4.0 kb EcoRI-DNA片段。通过珠孔插入注射吸管，注射几纳升DNA溶液（103拷贝/μ l）。注射卵的孵化频率与对照卵的孵化频率非常相似。在67条已发育到90天龄的鱼中，发现4条是转基因的，并且整合了1-5个拷贝/细胞的人生长激素基因构建体。在产卵后21-24 h注射的组中，含有外源DNA的鱼的数量最高（18条中有3条）。我们已经观察到，整合率低于哺乳动物。
The microinjection of the human growth hormone gene into the germinal disc and the development of 90-day live transgenic tilapia are reported. Three different spawns, treated 10 min, 21-24 h or 40-43 h after spawning, were used. For microinjection a linear 4.0 kb EcoRI-DNA fragment containing the mouse metallothionein-I promoter (mMT-I) fused to a structural gene coding for the human growth hormone (hGH) was used. The injection pipette was inserted through the micropyle and several nanoliters of DNA solution (103 copies/pl) were injected. Hatching frequencies of injected eggs were very similar to those of control eggs. Out of 67 fish which had developed to an age of 90 days, four were found to be transgenic and had integrated 1-5 copies/cell of the human growth hormone gene construct. The number of fish containing foreign DNA was highest (3 out of 18) in the group injected 21-24 h after spawning. We have observed that the integration rate is lower than in mammals.