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66, 1192 (1981) gpt delta Transgenic mouse Mutagen lEG10 DNA (80 copies/haploid in chromosome 17) Tissue in vitro packaging gpt delta mouse (C57BL/6J background) lEG10 phage Genomic DNA lEG10 chiC gpt Assay Spi- Assay 48 kb red, gam loxP loxP gpt cat Spi- Mutant Plaques 6-TGr mutant colonies Point Mutation Deletion NNK is a tobacco-specific nitrosamine that is carcinogenic in the lung of experimental animals NNK= nicotine-derived nitrosamino ketone O O N N C H 3 IARC :Group 2B (possible human carcinogen N 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanone Cytochrome P450 CYP2A Point Mutations DNA Adducts Cancer Lung O6-methylguanine O4-methylthymine Combined treatments of gpt delta mice with NNK and g-ray radiation Animal : gpt delta mouse, 8 mice (), 7 weeks old Saline NNK 2 mg/mouse/day x 4 (i.p.) Sacrifice Sacrifice 1.5 mGy/h 1.5 mGy/h 1.0 mGy/h 1.0 mGy/h 0.5 mGy/h 0.5 mGy/h 0 mGy/h 0 mGy/h (days) (days) Sakai et al., Int