Disney J E, Johnson K R, Banks D K, Thorgaard G H
Department of Zoology, Washington State University, Pullman 99164.
J Exp Zool. 1988 Dec;248(3):335-44. doi: 10.1002/jez.1402480312.
Transgenic rainbow trout (RT) were generated in earlier experiments by an in vivo modification of the chromosome-mediated gene transfer technique. This involved fertilizing albino RT eggs with gamma-irradiated brook (speckled) trout (ST) sperm and then heat shocking the eggs to induce second polar body retention. Striped pigment patterns were stable in six mosaic individuals between the ages of 7 and 14 months. We estimated the total ST DNA contribution in adult transgenic fish using DNA dot blot analyses. An average of 7% ST DNA was obtained in analyses of 11 transgenic individuals. Nine of 55 transgenic individuals examined expressed a ST-specific isozyme in adulthood; some were mosaic for foreign gene expression. Variable numbers of autonomous chromosome fragments were maintained in 10 adult transgenic individuals examined. Five transgenic RT were backcrossed to albino RT. Chromosome fragments were inherited by 29 of 31 backcross embryos examined. Additional active nucleolar organizer regions (NORs) were present on some fragments and may also have been integrated into some host chromosomes. This method may have numerous applications for basic and applied genetic research in fish.
在早期实验中,通过对染色体介导的基因转移技术进行体内改造,培育出了转基因虹鳟鱼(RT)。这包括用经γ射线照射的溪鳟(有斑点的鳟鱼,ST)精子使白化虹鳟鱼的卵子受精,然后对卵子进行热休克以诱导第二极体保留。在7至14个月大的6个嵌合体个体中,条纹色素模式是稳定的。我们使用DNA斑点印迹分析估计了成年转基因鱼中ST总DNA的贡献。在对11个转基因个体的分析中,平均获得了7%的ST DNA。在检查的55个转基因个体中,有9个在成年期表达了ST特异性同工酶;有些在异源基因表达方面是嵌合体。在检查的10个成年转基因个体中,维持了数量可变的自主染色体片段。将5个转基因虹鳟鱼与白化虹鳟鱼进行回交。在检查的31个回交胚胎中,有29个继承了染色体片段。一些片段上存在额外的活性核仁组织区(NORs),并且可能也已整合到一些宿主染色体中。这种方法可能在鱼类的基础和应用遗传研究中有许多应用。