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Transgenic zebrafish for detecting mutations caused by compounds in aquatic environments

Nature Biotechnology volume 18, pages 62–65 (2000)Cite this article

Abstract

We have established a transgenic zebrafish line carrying a shuttle vector plasmid (pML4) for detecting mutagens in aquatic environments. The plasmid contains the rpsL gene of Escherichia coli as a mutational target gene, and the kanamycin-resistance gene for recovering the plasmid from the chromosomal DNA. To evaluate the system, we treated embryos of the transgenic fish with N-ethyl-N-nitrosourea (ENU), which induces a dose-dependent increase in the mutation frequency of the target gene. The mutation spectrum was consistent with the proposed mechanism of ENU mutagenesis. Similarly, treating the embryos with benzo[a]pyrene or 2-amino-3,8-dimethylimidazo[4,5- f]quinoxaline, which are found in naturally polluted water, significantly increased the frequency of mutations in the target gene.

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Figure 1: Estimation of the copy number of the integrated shuttle vector plasmid by Southern blot analysis.
Figure 2: Mutant frequency of the transgene in embryos after treatment with B[ a]P and MeIQx.
Figure 3

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Acknowledgements

We are grateful to Hisaji Maki and Mutsuo Sekiguchi for providing us with the pML4 plasmid, and to Yoichi Gondo and Motoya Katsuki for discussion on the rescue of the plasmid. We also thank Chizuko Iijima, Keiko Miki, and Setsuko Kunimoto for technical assistance and for maintaining fish, and Hiroyuki Haraguchi for sequence analysis. This work was supported in part by a grant from Sumitomo Foundation.

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Author notes

  1. Hiroyuki Takeda

    Present address: Division of Early Embryogenesis, National Institute of Genetics, Mishima, 411-8540, Japan

Authors and Affiliations

  1. Environmental Health Science Division, National Institute for Environmental Studies, Onogawa, Tsukuba, 305-0053, Japan

    Kimiko Amanuma & Yasunobu Aoki

  2. Molecular Cell Science Laboratory, Tsukuba Life Science Center, The Institute of Physical and Chemical Research (RIKEN), Koyadai, Tsukuba, 305-0074, Ibaraki, Japan

    Hiroshi Amanuma

  3. Department of Molecular Biology, School of Science, Nagoya University, Chikusaku, Nagoya, 464-8602, Japan

    Hiroyuki Takeda

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Correspondence to Kimiko Amanuma.

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Amanuma, K., Takeda, H., Amanuma, H. et al. Transgenic zebrafish for detecting mutations caused by compounds in aquatic environments. Nat Biotechnol 18, 62–65 (2000). https://doi.org/10.1038/71938

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