Tissue Regeneration Project, Hiroshima Prefecture Collaboration of Regional Entities for the Advancement of Technological Excellence, Japan Science and Technology Corporation, Hiroshima Prefecture Institute of Industrial Science and Technology, Higashihiroshima, Hiroshima 739-0046, Japan; Laboratory of Developmental Biology, Department of Biological Science, Graduate School of Science, Hiroshima University, 1-3-1 Kagamiyama, Higashihiroshima, Hiroshima 739-8526, Japan; ProPhoenix Company Ltd., Higashihiroshima, Hiroshima 739-0046, Japan; Towa-Kagaku Corporation Ltd., Hiroshima, Hiroshima 730-0841, Japan.
Environ Toxicol Pharmacol. 2003 Apr;13(3):153-9. doi: 10.1016/S1382-6689(03)00003-6.
We generated germ line-transgenic Xenopus laevis that monitors environmental heavy metal ions. Sperm nuclei were transduced with cDNA of enhanced green fluorescent protein (EGFP) driven by murine metallothionein-1 gene promoters and were microinjected into unfertilized eggs. The eggs developed to sexually matured adults. The transgenic tadpoles at the premetamorphic stage were reared in water containing Zn(2+) and Cd(2+) separately at the concentrations of 0.38-1.52 and 0.09-0.44 μM, respectively. These animals responded to Zn(2+) at as low as 0.38 μM and Cd(2+) at as low as 0.44 μM. The level of EGFP fluorescence emitted by tadpoles increased as the concentration increased up to 1.52 μM and the exposure time prolonged up to 120 h. The fluorescent response was much more sensitive to Cd(2+) than to Zn(2+). We concluded that these transgenic tadpoles are useful as an animal indicator of environmental heavy metal ions.
我们生成了可监测环境重金属离子的品系转化非洲爪蟾。精子细胞核被增强型绿色荧光蛋白（EGFP）的 cDNA 转导，该 cDNA 由鼠金属硫蛋白-1 基因启动子驱动，并被微注射到未受精的卵中。这些卵发育为性成熟的成体。处于前变态期的转基因蝌蚪在含有 Zn（2+）和 Cd（2+）的水中分别以 0.38-1.52 和 0.09-0.44 μM 的浓度进行培养。这些动物对低至 0.38 μM 的 Zn（2+）和低至 0.44 μM 的 Cd（2+）有反应。随着浓度增加到 1.52 μM 和暴露时间延长到 120 h，蝌蚪发出的 EGFP 荧光水平增加。荧光反应对 Cd（2+）比对 Zn（2+）更为敏感。我们得出结论，这些转基因蝌蚪可用作环境重金属离子的动物指示剂。
