Graduate Institute of Engineering Technology-Doctoral, National Taipei University of Technology, 1 Chung-Hsiao E. Rd., Sec. 3, Taipei, 10608, Taiwan.
Fish Physiol Biochem. 2013 Dec;39(6):1525-39. doi: 10.1007/s10695-013-9806-6. Epub 2013 May 14.
One cannot seek permission to market transgenic fish mainly because there is no field test or any basic research on technological developments for evaluating their biosafety. Infertility is a necessary adjunct to exploiting transgenic fish unless completely secure land-locked facilities are available. In this study, we report the generation of a Cre transgenic zebrafish line using a cytomegalovirus promoter. We also produced fish carrying the Bax1 and Bax2 plasmids; these genes were separated by two loxP sites under a zona pellucida C promoter or were driven by an anti-Müllerian hormone promoter. We inserted a red fluorescent protein gene between the two loxP sites. After obtaining transgenic lines with the two transgenic fish crossed with each other (Cre transgenic zebrafish x loxP transgenic zebrafish), the floxed DNA was found to be specifically eliminated from the female or male zebrafish, and apoptosis gene expressions caused ovarian and testicular growth cessation and degeneration. Overexpression of the Bax1 and Bax2 genes caused various expression levels of apoptosis-related genes. Accordingly, this transgenic zebrafish model system provides a method to produce infertile fish and may be useful for application to genetically modified fish.
由于缺乏对转基因鱼进行田间试验或任何基本技术发展评估其生物安全性的研究，因此不能寻求许可来推广转基因鱼。除非有完全安全的陆基设施，否则不育是开发转基因鱼的必要手段。在这项研究中，我们使用巨细胞病毒启动子报告了 Cre 转基因斑马鱼系的产生。我们还产生了携带 Bax1 和 Bax2 质粒的鱼；这些基因在透明带 C 启动子下或在抗苗勒氏管激素启动子的驱动下被两个 loxP 位点隔开。我们在两个 loxP 位点之间插入了一个红色荧光蛋白基因。通过将两条转基因鱼（Cre 转基因斑马鱼 x loxP 转基因斑马鱼）相互杂交获得转基因系后，发现 floxed DNA 被特异性地从雌性或雄性斑马鱼中消除，凋亡基因表达导致卵巢和睾丸生长停止和退化。Bax1 和 Bax2 基因的过表达导致凋亡相关基因的不同表达水平。因此，这种转基因斑马鱼模型系统提供了一种产生不育鱼的方法，可能对遗传修饰鱼的应用有用。
