Peters K G, Rao P S, Bell B S, Kindman L A
Department of Medicine, Duke University Medical Center, Durham, North Carolina 27710, USA.
Dev Biol. 1995 Sep;171(1):252-7. doi: 10.1006/dbio.1995.1276.
The recent development of transgenic technology in zebrafish has opened an exciting new avenue in which to explore vertebrate development. However, as in other species, the inability to easily identify live transgenic fish severely limits the potential of this promising technology. To determine whether the recently described green fluorescent protein (GFP) might provide a convenient live staining method in zebrafish, we constructed a glutathione S-transferase/GFP fusion protein (GST-GFP). GST-GFP cRNA, when injected into individual blastomeres of early zebrafish embryos, resulted in the rapid development (3 hr) of easily detectable green fluorescence which persisted for up to 4 days. GFP fluorescence was restricted to progeny of the injected cell and appeared to have no adverse effects on embryonic development despite widespread expression. Our findings demonstrate that GFP fusion proteins will provide a simple yet powerful means of monitoring production of heterologous proteins in live zebrafish.
斑马鱼转基因技术的最新进展为探索脊椎动物发育开辟了一条令人兴奋的新途径。然而，与其他物种一样，难以轻松鉴定活的转基因鱼严重限制了这项有前途技术的潜力。为了确定最近描述的绿色荧光蛋白（GFP）是否能为斑马鱼提供一种方便的活体染色方法，我们构建了一种谷胱甘肽S - 转移酶/GFP融合蛋白（GST - GFP）。当将GST - GFP cRNA注射到斑马鱼早期胚胎的单个卵裂球中时，可快速（3小时）产生易于检测的绿色荧光，这种荧光可持续长达4天。GFP荧光仅限于注射细胞的后代，尽管广泛表达，但似乎对胚胎发育没有不利影响。我们的研究结果表明，GFP融合蛋白将为监测活斑马鱼中异源蛋白的产生提供一种简单而强大的方法。
