1997 Fiscal Year Annual Research Report

遺伝子導入マウスを用いた組織別突然変異の鋭敏簡便な検出とDNA変化の解析

Research Project

Project/Area Number	07554043
Research Institution	TOKAI UNIVERSITY
Principal Investigator	権藤洋一東海大学, 総合医学研究所, 助教授 (40225678)
Co-Investigator(Kenkyū-buntansha)	美野輪治 (財)癌研究会癌研究所, 細胞生物部, 研究員 (00181967) 中津可道大阪大学, 細胞生体工学センター, 助手 (00207820) 丹羽太貫京都大学, 放射線生物研究センター, 教授 (80093293) 真木寿治奈良先端科学技術大学院大学, バイオサイエンス研究科, 教授 (20199649) 祖父尼俊雄国立医薬品食品衛生研究所, 変異遺伝部, 部長(研究職) (20132889)
Keywords	トランスジェニックマウス / 突然変異 / rpsL / 発がんリスク評価 / 遺伝毒性検定 / 環境変異原 / 変異原性試験 / シャトルベクター
Research Abstract	突然変異を容易に検出できるシャトルベクターを導入したrpsLトランスジェニックマウスを開発し、自然発生突然変異や環境変異原のもつ遺伝子毒性を、生きたマウス個体の発生段階別、各臓器別に、また、遺伝子修復酵素を欠損するマウスと交配することも含めて、検定できる系を確立している。平成9年度は、権藤が開発した新しいシャトルベクターを導入したトランスジェニックマウスを用いたデータも蓄積し、1)バックグランド突然変異体頻度が1×10^<-5>に下がる、2)検定効率が3倍になる、3)メチル化されたシャトルベクターも効率よく検定できる、4)生じる塩基変化は基本的に従来の系と同じである、ということが明らかになった(権藤、中村、若林)。真木は、大腸菌系におけるモニター遺伝子そのものの改良を行いエラーを起こしやすい配列を同義置換によって変えたことろバックグランドが低くなることを確認した。祖父尼は従来の系に加えさらにラムダベクター内のcII遺伝子を利用したポジティブ選択系を開発した。丹羽は放射線投与をマウスミニサテライトに加え、Droshphilaのwhite遺伝子座においても検討しこれまでの結果を指示するデータを得た。中津および美野輪は、新しいトランスジェニックマウスと、XPA欠損マウスおよびMLH1遺伝子欠損マウスとのF2マウスが得られ、それぞれ突然変異解析を始めた。さらに、rpsLトランスジェニックマウスを用いたDNA変化の解析の結果、生物個体に遺伝毒性がもたらされているときには塩基置換が、細胞毒性がもたらされているときには1塩基欠失によるフレームシフト突然変異がモニター遺伝子であるrpsL上に現われることが示唆され、今後、薬剤の発がん性と抗がん性を検証する場合、有効な指標となる可能性が示された。

Research Products

(19 results)

All Other

All Publications (19 results)

[Publications] Y.Gondo: "Analysis of genetic instabilities of RS447 megasatellite DNA region in human 4p15." American Journal of Human Genetics. 61(4). A391 (1997)
[Publications] T.Suzuki: "Ethyl nitrosourea and methyl methanesulfonate mutagenicity in sperm and testicular germ cells of lacZ transgenic mice(Muta TMMouse)." Mutation Research. 388. 155-163 (1997)
[Publications] H.Okonogi: "Agreement of mutational characteristics of heterocyclic amines in lacI of the Big Blue mouse with those in tumor related genes in rodents." Carcinogenesis. 18. 745-748 (1997)
[Publications] A.Nishikawa: "Comparative study on organ-specificity of tumorigenicity,mutagenicity and cell proliferative activity induced by dimethylnitrosamine in Big Blue mice." Cancer Letters. 117. 143-147 (1997)
[Publications] T.Ono: "X-ray-and ultraviolet-radiation-induced mutations in Muta TMMouse." Radiation Research. 148. 123-128 (1997)
[Publications] N.Kotomura: "Transcriptional regulatin by competition between ELP isoforms and nuclear receptors." Biochemical and Biophysical Research Communication. 230. 407-412 (1997)
[Publications] S.Tashiro: "Nuclear proteins binding to the recombination hotspot region of the retinoic acid receptor alpha gene." Leukemia. 11. 285-286 (1997)
[Publications] Y.Abe: "A defect in cell to celladhesion via integrin-fibronectin interactins observed in a highly metastatic tumor cell line." Japanese Journal of Cancer Research. 88. 64-71 (1997)
[Publications] K.Shimokado: "Agreement of mutational characteristics of heterocyclic amines in lacI of the Big Blue mouse with those in tumor related genes in rodents." Japanese Journal of Cancer Research. (in press). (1998)
[Publications] Y.Enokido: "Loss of the Xeroderma Pigmentosum Group A Gene(XPA)Enhances Apoptosis of Cultured Cerebellar Neurons Induced by UV but Not by Low-K+Medium." Jounal of Neurochemistry. 69. 246-251 (1997)
[Publications] J.-P.Yeo: "RPA2,a gene for the 32 kDa subunit of replication protein A on chromosome 1p35-36,is not mutated in patients with familial melanoma linked to chromosome 1p36." Melanoma Research. 8. 1-6 (1997)
[Publications] S.Takeuchi: "Strand specificity and absence of hotspots for p53 mutations in UVB-induced skin tumors of XPA deficient mice." Cancer Research. (in press). (1998)
[Publications] H.Beppu: "cDNA Cloning and Genomic Organization of the Mouse BMP Type II Receptore." Biochemical and Biophysical Research Communication. 235. 499-504 (1997)
[Publications] J.Aruga: "Mouse Zic 1 is involved in cerebellar development." Journal of Neuroscience. 18. 284-293 (1998)
[Publications] T.Matsuzaki: "Superovulation induction in the House Musk Shrew(Suncus murinus)." Experimental Animals. 46. 183-189 (1997)
[Publications] T.Matsuzaki: "The period of ovulation and presence of the first polar body of ova ovulated in the House Musk Shrew(Suncus murinus)." Experimental Animals. 46. 291-296 (1997)
[Publications] N.Shinohara: "Prevention of autoantibody production in lpr/lpr mice by transgenic expression of Fas on B cells." B Lymphocytes and Autoimmunity. 815. 489-491 (1997)
[Publications] K.Koera: "K-Ras is essential for the development of the mouse embryo." Oncogene. 15. 1151-1159 (1997)
[Publications] 真木寿治: "「シリーズ分子生物学第2巻遺伝子工学」大腸菌での遺伝子操作." 朝倉書店, 21 (1997)

1997 Fiscal Year Annual Research Report

遺伝子導入マウスを用いた組織別突然変異の鋭敏簡便な検出とDNA変化の解析

Principal Investigator

権藤 洋一 東海大学, 総合医学研究所, 助教授 (40225678)

Research Products

[Publications] Y.Gondo: "Analysis of genetic instabilities of RS447 megasatellite DNA region in human 4p15." American Journal of Human Genetics. 61(4). A391 (1997)

[Publications] T.Suzuki: "Ethyl nitrosourea and methyl methanesulfonate mutagenicity in sperm and testicular germ cells of lacZ transgenic mice(Muta TMMouse)." Mutation Research. 388. 155-163 (1997)

[Publications] H.Okonogi: "Agreement of mutational characteristics of heterocyclic amines in lacI of the Big Blue mouse with those in tumor related genes in rodents." Carcinogenesis. 18. 745-748 (1997)

[Publications] A.Nishikawa: "Comparative study on organ-specificity of tumorigenicity,mutagenicity and cell proliferative activity induced by dimethylnitrosamine in Big Blue mice." Cancer Letters. 117. 143-147 (1997)

[Publications] T.Ono: "X-ray-and ultraviolet-radiation-induced mutations in Muta TMMouse." Radiation Research. 148. 123-128 (1997)

[Publications] N.Kotomura: "Transcriptional regulatin by competition between ELP isoforms and nuclear receptors." Biochemical and Biophysical Research Communication. 230. 407-412 (1997)

[Publications] S.Tashiro: "Nuclear proteins binding to the recombination hotspot region of the retinoic acid receptor alpha gene." Leukemia. 11. 285-286 (1997)

[Publications] Y.Abe: "A defect in cell to celladhesion via integrin-fibronectin interactins observed in a highly metastatic tumor cell line." Japanese Journal of Cancer Research. 88. 64-71 (1997)

[Publications] K.Shimokado: "Agreement of mutational characteristics of heterocyclic amines in lacI of the Big Blue mouse with those in tumor related genes in rodents." Japanese Journal of Cancer Research. (in press). (1998)

[Publications] Y.Enokido: "Loss of the Xeroderma Pigmentosum Group A Gene(XPA)Enhances Apoptosis of Cultured Cerebellar Neurons Induced by UV but Not by Low-K+Medium." Jounal of Neurochemistry. 69. 246-251 (1997)

[Publications] J.-P.Yeo: "RPA2,a gene for the 32 kDa subunit of replication protein A on chromosome 1p35-36,is not mutated in patients with familial melanoma linked to chromosome 1p36." Melanoma Research. 8. 1-6 (1997)

[Publications] S.Takeuchi: "Strand specificity and absence of hotspots for p53 mutations in UVB-induced skin tumors of XPA deficient mice." Cancer Research. (in press). (1998)

[Publications] H.Beppu: "cDNA Cloning and Genomic Organization of the Mouse BMP Type II Receptore." Biochemical and Biophysical Research Communication. 235. 499-504 (1997)

[Publications] J.Aruga: "Mouse Zic 1 is involved in cerebellar development." Journal of Neuroscience. 18. 284-293 (1998)

[Publications] T.Matsuzaki: "Superovulation induction in the House Musk Shrew(Suncus murinus)." Experimental Animals. 46. 183-189 (1997)

[Publications] T.Matsuzaki: "The period of ovulation and presence of the first polar body of ova ovulated in the House Musk Shrew(Suncus murinus)." Experimental Animals. 46. 291-296 (1997)

[Publications] N.Shinohara: "Prevention of autoantibody production in lpr/lpr mice by transgenic expression of Fas on B cells." B Lymphocytes and Autoimmunity. 815. 489-491 (1997)

[Publications] K.Koera: "K-Ras is essential for the development of the mouse embryo." Oncogene. 15. 1151-1159 (1997)

[Publications] 真木 寿治: "「シリーズ分子生物学 第2巻 遺伝子工学」大腸菌での遺伝子操作." 朝倉書店, 21 (1997)

権藤洋一東海大学, 総合医学研究所, 助教授 (40225678)

[Publications] 真木寿治: "「シリーズ分子生物学第2巻遺伝子工学」大腸菌での遺伝子操作." 朝倉書店, 21 (1997)