Other transgenic mutation assays: A new transgenic mouse mutagenesis test system using Spi− and 6-thioguanine selections
1996; Wiley; Volume: 28; Issue: 4 Linguagem: Inglês
10.1002/(sici)1098-2280(1996)28
ISSN1098-2280
AutoresT. Nohmi, Masaya Katoh, Hiroshi Suzuki, M. Matsui, Masami Yamada, Masao Watanabe, Makoto Suzuki, N. Horiya, Otoya Ueda, Tohru Shibuya, H. Ikeda, Toshio Sofuni,
Tópico(s)Advanced biosensing and bioanalysis techniques
ResumoEnvironmental and Molecular MutagenesisVolume 28, Issue 4 p. 465-470 Original Article Other transgenic mutation assays: A new transgenic mouse mutagenesis test system using Spi− and 6-thioguanine selections T. Nohmi, Corresponding Author T. Nohmi Division of Genetics and Mutagenesis, National Institute of Health Sciences, Setagaya-ku, Tokyo, JapanDivision of Genetics and Mutagenesis, National Institute of Health Sciences, 1-18-1 Kamiyoga, Setagaya-ku, Tokyo 158, JapanSearch for more papers by this authorM. Katoh, M. Katoh Faculty of Odontology, University of Chile, Santiago, ChileSearch for more papers by this authorH. Suzuki, H. Suzuki Exploratory Research Laboratory, Fuji Gotemba Research Laboratory, Chugai Pharmaceutical Co., Ltd., Gotemba, Shizuoka, JapanSearch for more papers by this authorM. Matsui, M. Matsui Division of Genetics and Mutagenesis, National Institute of Health Sciences, Setagaya-ku, Tokyo, JapanSearch for more papers by this authorM. Yamada, M. Yamada Division of Genetics and Mutagenesis, National Institute of Health Sciences, Setagaya-ku, Tokyo, JapanSearch for more papers by this authorM. Watanabe, M. Watanabe Division of Genetics and Mutagenesis, National Institute of Health Sciences, Setagaya-ku, Tokyo, Japan Biochemistry Division, National Cancer Center Research Institute, Chuoku, Tokyo, JapanSearch for more papers by this authorM. Suzuki, M. Suzuki Division of Genetics and Mutagenesis, National Institute of Health Sciences, Setagaya-ku, Tokyo, JapanSearch for more papers by this authorN. Horiya, N. Horiya Laboratory of Genetic Toxicology, Hatano Research Institute, Hadano, Kanagawa, JapanSearch for more papers by this authorO. Ueda, O. Ueda Exploratory Research Laboratory, Fuji Gotemba Research Laboratory, Chugai Pharmaceutical Co., Ltd., Gotemba, Shizuoka, JapanSearch for more papers by this authorT. Shibuya, T. Shibuya Laboratory of Genetic Toxicology, Hatano Research Institute, Hadano, Kanagawa, JapanSearch for more papers by this authorH. Ikeda, H. Ikeda Department of Molecular Biology, The Institute of Medical Science, The University of Tokyo, Tokyo, JapanSearch for more papers by this authorT. Sofuni, T. Sofuni Laboratory of Genetic Toxicology, Hatano Research Institute, Hadano, Kanagawa, JapanSearch for more papers by this author T. Nohmi, Corresponding Author T. Nohmi Division of Genetics and Mutagenesis, National Institute of Health Sciences, Setagaya-ku, Tokyo, JapanDivision of Genetics and Mutagenesis, National Institute of Health Sciences, 1-18-1 Kamiyoga, Setagaya-ku, Tokyo 158, JapanSearch for more papers by this authorM. Katoh, M. Katoh Faculty of Odontology, University of Chile, Santiago, ChileSearch for more papers by this authorH. Suzuki, H. Suzuki Exploratory Research Laboratory, Fuji Gotemba Research Laboratory, Chugai Pharmaceutical Co., Ltd., Gotemba, Shizuoka, JapanSearch for more papers by this authorM. Matsui, M. Matsui Division of Genetics and Mutagenesis, National Institute of Health Sciences, Setagaya-ku, Tokyo, JapanSearch for more papers by this authorM. Yamada, M. Yamada Division of Genetics and Mutagenesis, National Institute of Health Sciences, Setagaya-ku, Tokyo, JapanSearch for more papers by this authorM. Watanabe, M. Watanabe Division of Genetics and Mutagenesis, National Institute of Health Sciences, Setagaya-ku, Tokyo, Japan Biochemistry Division, National Cancer Center Research Institute, Chuoku, Tokyo, JapanSearch for more papers by this authorM. Suzuki, M. Suzuki Division of Genetics and Mutagenesis, National Institute of Health Sciences, Setagaya-ku, Tokyo, JapanSearch for more papers by this authorN. Horiya, N. Horiya Laboratory of Genetic Toxicology, Hatano Research Institute, Hadano, Kanagawa, JapanSearch for more papers by this authorO. Ueda, O. Ueda Exploratory Research Laboratory, Fuji Gotemba Research Laboratory, Chugai Pharmaceutical Co., Ltd., Gotemba, Shizuoka, JapanSearch for more papers by this authorT. Shibuya, T. Shibuya Laboratory of Genetic Toxicology, Hatano Research Institute, Hadano, Kanagawa, JapanSearch for more papers by this authorH. Ikeda, H. Ikeda Department of Molecular Biology, The Institute of Medical Science, The University of Tokyo, Tokyo, JapanSearch for more papers by this authorT. Sofuni, T. Sofuni Laboratory of Genetic Toxicology, Hatano Research Institute, Hadano, Kanagawa, JapanSearch for more papers by this author First published: 1996 https://doi.org/10.1002/(SICI)1098-2280(1996)28:4 3.0.CO;2-CCitations: 184AboutPDF ToolsRequest permissionExport citationAdd to favoritesTrack citation ShareShare Give accessShare full text accessShare full-text accessPlease review our Terms and Conditions of Use and check box below to share full-text version of article.I have read and accept the Wiley Online Library Terms and Conditions of UseShareable LinkUse the link below to share a full-text version of this article with your friends and colleagues. Learn more.Copy URL Share a linkShare onFacebookTwitterLinked InRedditWechat Abstract A new transgenic mouse mutagenesis test system has been developed for the efficient detection of point mutations and deletion mutations in vivo. The mice carry lambda EG10 DNA as a transgene. When the rescued phages are infected into Escherichia coli YG6020-expressing Cre recombinase, the phage DNA is converted into plasmid pYG142 carrying the chloramphenicol-resistance gene and the gpt gene of E. coli. The gpt mutants can be positively detected as colonies arising on plates containing chlaramphenicol and 6-thioguanine. The EG10 DNA carries a chi site along with the red and gam genes so that the wild-type phages display Spi+ (sensitive to P2 interference) phenotype. Mutant phages lacking both red and gam genes can be positively detected as plaques that grow in P2 lysogens of E. coli. These mutant phages are called lambda Spi−. The spontaneous gpt mutation frequencies of five independent transgenic lines were 1.7 to 3.3 × 10−5 in bone marrow. When the mice were treated with ethylnitrosourea (single i.p. treatments with 150 mg/kg body weight; killed 7 days after the treatments), mutation frequencies were increased four- to sevenfold over the background in bone marrow. The average rescue efficiencies were more than 200,000 chloramphenicol-resistant colonies per 7.5 μg bone marrow DNA per packaging reaction. In contrast to gpt mutation frequencies, spontaneous Spi− mutation frequencies were 1.4 × 10−6 and 1.1 × 10−6 in bone marrow and sperm, respectively. No spontaneous Spi− mutants have been detected so far in spleen, although 930,000 phages rescued from untreated mice were screened. In gamma-ray-treated animals, however, induction of Spi− mutations was clearly observed in spleen, at frequencies of 1.4 × 10−5 (5 Gy), 1.2 × 10−5 (10 Gy), and 2.0 × 10−5 (50 Gy). These results suggest that the new transgenic mouse "gpt delta" could be useful for the efficient detection of point mutations and deletion mutations in vivo. © 1996 Wiley-Liss Inc. Citing Literature Volume28, Issue41996Pages 465-470 RelatedInformation
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