1995 Fiscal Year Annual Research Report

標的遺伝子組換えによる突然変異体の作成とその生体機能の研究

Research Project

Project/Area Number	05275102
Research Institution	The University of Tokyo
Principal Investigator	勝木元也東京大学, 医科学研究所, 教授 (20051732)
Co-Investigator(Kenkyū-buntansha)	前田秀一郎山梨医科大学, 生化学第一, 教授 (10117244) 村松喬名古屋大学, 医学部, 教授 (00030891) 林崎良英理化学研究所, ライフ・筑波研究センター, 主任研究員 (70192705) 金田安史大阪大学, 細胞生体工学センター, 助教授 (10177537) 糸原重美京都大学, ウィルス研究所, 助教授 (60252524)
Keywords	標的遺伝子組換え / ジーンターゲッティング / 中枢神経系 / 免疫系 / 形態形成 / グルタミン酸受容体 / ドーパミン受容体 / 突然変異
Research Abstract	本研究は標的遺伝子組換え法(ジーンターゲッティング法)を用いて、中枢神経系、免疫系、形態形成等に関与する遺伝子を計画的に破壊し、それぞれの遺伝子の生体での機能を解析することを目的としている。計画班員は、きわめて緊密に連携し、独立を保ちながら、多種の遺伝子の突然変異体の作成に成功した。本年度までに本研究班によって作成された突然変異マウスを列挙すれば90種を越えるに至った。グルタミン酸受容体(NMDAタイプ5種、およびmGluR4種)、ドーパミン受容体、セロトニン受容体、リアノジン受容体、IP3受容体、H-ras、N-ras、K-ras、rac1、rac2、などである。詳細は、進捗状況報告および研究成果報告書にまとめてある。それぞれの遺伝子破壊によって認められる生体の特徴は、細胞レベルやタンパク質化学レベルの既存の成果から容易に予測されものは少なく、それぞれの遺伝子産物が多くの生体機能に関わっていることが示された。また、行動異常や記憶や学習の実験などにより高次機能の遺伝子発現との関係が検討された。本年度はジーンターゲッティングマウスが次々に作成され、来年度にかけてその解析の結果が続々と報告されるものと期待される。

Research Products

(48 results)

All Other

All Publications (48 results)

[Publications] Yamaguchi, H., et al.: "Dopamine D2 receptor plays a critical role in cell proliferation and proopiomelanocortin expression in the pituitary." Genes to Cells. in press.
[Publications] Gondo, Y., et al.: "A novel positive detection system of in vivo mutations in rpsL(strA)transgenic mice." Mutation Res. in press.
[Publications] Masu, M., et al.: "Specific deficit of the ON response in visual transmission by targeted disruption of the mGluR6 gene." Cell. 80. 757-765 (1995)
[Publications] Shimano, H., et al.: "Inhibition of diet-induced atheroma formation in transgenic mice expressing apolipoprotein E in the arterial wall." J Clin Invest. 95. 469-476 (1995)
[Publications] Honda, H., et al.: "Increased tyrosine-phosphorylation of 55KDa proteins in beta-actin/Tec transgenic mice." Biochem Biophys Res Commun. 206. 287-293 (1995)
[Publications] Tamura, H., et al.: "Opaque eyes developed in transgenic mice with T-cell receptor δ gene." Invest Ophthalmol Vis Sci. 36. 467-477 (1995)
[Publications] Komano, H., et al: "Homeostatic regulation of intestinal epithelia by intraepithelial γδ T cells." Proc Natl Acad Sci USA. 92. 6147-6151 (1995)
[Publications] Gomi, H., et al.: "Mice devoid of the glial fibrillary acidic protein develop normally and are susceptible to scrapie prions." Neuron. 14. 29-41 (1995)
[Publications] Emoto, M., et al.: "Mice dificient in γδ T cells are resistant to lethal infection with Salmonella choleraesuis." Infect Immun. 63. 3736-3738 (1995)
[Publications] Nishimura, H., et al.: "The role of γδ T cells in priming macrophages to produce tumor necrosis factor-α." Eur Immunol. 25. 1465-1468 (1995)
[Publications] Nakane, H., et al.: "High incidence of UVB-or chemical carcinogen-induced skin tumorigenesis in mice lacking the xerodema pigmentosum group A(XPA)gene." Nature. in press.
[Publications] Kaneda, Y., et al.: "The recycling of synaptic vesicle membrane components. “Protein Kinesis: The Dynamics of Protein Trafficking and Stability"" Cold Spring Harbor Symp. Quant. Biol. 60(in press).
[Publications] Mann, M. J., et al.: "Genetic engineering of vein grafts resistant to atherosclerosis." Proc Natl Acad Sci USA. 92. 4502-4506 (1995)
[Publications] von der Leyen, H. E., et al.: "Gene therapy inhibiting neointimal vascular lesion: In vivo transfer of endothelial cell nitric oxide synthase gene." Proc Natl Acad Sci USA. 92. 1137-1141 (1995)
[Publications] Kaneda. Y., et al.: "Increased wxpression of DNA cointroduced with nuckear protein in adult rat liver." J Mol Med. 73. 289-297 (1995)
[Publications] Mizuki, N., et al.: "Isolation of cDNA and genomic clones of a human Ras-related GTP-binding protein gene and its choromosomal localization to the long arm of choromosome 7, 7q36." Genomics. in press.
[Publications] Tsai, HJ., et al.: "Initiation of the transgenic lacZ gene expression in medaka(Oryzias latipes)embryos." Mol Mar Biol Biotechnol. 4. 1-9 (1995)
[Publications] Naruse, T., et al.: "Cloned PLT cells recognize the forth,fifth and sixth hypervariable regions at amino acid positions 65 to 87 of DPB1 molecule." Human Immunol. 42. 123-130 (1995)
[Publications] Kajiwara, K., et al.: "Cloning and characterization of pentylenetetrazol-related cDNA, PTZ-17." Brain Res. 671. 170-174 (1995)
[Publications] Cai, J-P., et al.: "cDNA and genomic sequences for rat 8-oxo-dGTPase that prevents occurrence of spontaneous mutations due to oxidation of guanine nucleotides." Carcinogenesis. 16. 2343-2350 (1995)
[Publications] Kakuma, T., et al.: "Mouse MTH1 protein with 8-Oxo-7,8-dihydro-2′-deoxyguanosine5′-triphatase activity that prevents transversion mutation." J Biological Chem. 270. 2542-2548 (1995)
[Publications] Rancourt, D. E., et al.: "Genetic interaction between hoxb-5 and hoxb-6 is revealed by nonallelic noncomplementation." Genes Dev. 9. 108-122 (1995)
[Publications] Kuromitsu, J., et al.: "Programmed alterations of DNA methylation at CpG islands during blastformation of peripheral blood lymphocytes." DNA Res. in press.
[Publications] Plass, C., et al.: "Comparative analysis of mouse NotI Iinking clones with mouse and human genomic sequences and transcripts." DNA Res. 2. 27-35 (1995)
[Publications] Hirotsune, S., et al.: "The reeler gene encodes a protein with EGF-like motif expressed by pioneer neurons." Nature Genetics. 10. 77-83 (1995)
[Publications] Latham, K., et al.: "Stage-specific induction and regulation by genomic imprinitng of mouse U2bpaf-rs gene during preimplantation development." Developmental Biology. 168. 670-676 (1995)
[Publications] Okazaki, Y., et al.: "Direct detection and isolation of RLGS spot DNA marker tightly linked to a specific trait using RLGS spot-bombing method." Proc Natl Acad Sci USA. 92. 5610-5614 (1995)
[Publications] Ohsumi, T., et al.: "Loss of heterozygosity in chromosomes 1, 5, 7and13 in mouse hepatoma detected by systematic genome-wide scanning RLGS genetic map." Biochem Biophys Res Commun. 212. 632-639 (1995)
[Publications] Kalcheva, I., et al.: "Comparative mapping of the imprinted U2afbpL gene on mouse chromosome11 and human chromosome5." Cytogenet Cell grenet. 68. 19-24 (1995)
[Publications] Okazaki, Y, et al.: "Amultiplex procduction system of RLGS gel-A new version of restriction landmark genomic scanning method(RLGS Ver1, 8)." Electrophoresis. 16. 197-202 (1995)
[Publications] Ohsumi, T, et al.: "RLGS spot cloning method." Electrophoresis. 16. 203-209 (1995)
[Publications] Shibata, H., et al.: "Systematic screening of endogenous imprinted genes using RLGS and identification of a new imprinted gene U2af binding protein related sequence." Electrophoresis. 16. 210-217 (1995)
[Publications] Watanabe, S., et al.: "Accessibilitey to tissue-specific genes from methylantion profiles of mouse brain genomic DNA." Electrophoresis. 16. 218-226 (1995)
[Publications] Okuizumi, H., et al.: "Genetic mapping of restriction landmark senomic scanning(RLGS)loci in the mouse." Electrophoresis. 16. 233-240 (1995)
[Publications] Okuizumi, H., et al.: "A single gel analysis of 573 dominant and codominant restriction landmark genomic scanning(RLGS)loci in interspecific backcross progeny." Electrophoresis. 16. 253-260 (1995)
[Publications] Horie, K., et al.: "A replacement vector us ed to introduce subtle mutaions into mouse genes." Gene. in press.
[Publications] Kato, G. et al.: "Novel phosphory lation at a mitotic site, serine 75,in human pp60^<c-src> from unsynchronized human tumor cells having a spherical morphology." Biochem Biophys Res Commun. 216. 619-629 (1995)
[Publications] Wei, S., et al.: "Studies on the metabolism of retinol and retinol-binding protein in transthyretin-deficient mice produced by homologous recombi-nation." J Biol Chem. 270. 866-870 (1995)
[Publications] Matsuo. I., et al.: "Mouse Ot×2 functions in the formation and patteming of rostral head." Gene Dev. 9. 2646-2658 (1995)
[Publications] Miyauchi, T., et al.: "Structure of the mouse basigin gene,a unique member of the immunoglobulin superfamily." J Biochem. in press.
[Publications] Masuda, K., et al.: "Functional rescue of photoreceptos from the damaging effects of constant light by survival-promoting factors in the rat." Invest Ophtalmol Vis Sci. in press.
[Publications] Obama, H., et al.: "A transgenic mouse line with α-1,3/4 fucosyltransferase cDNA:production and characteristics." Glycoconjugate J. 12. 795-801 (1995)
[Publications] Yu, S., et al.: "cDNA cloning and sequence analysis of a novel calcium binding protein with oligoproline motif" Cell Struct Funct. 20. 263-268 (1995)
[Publications] Kojima, S., et al.: "Midkine enhances fibrinolytic activity of bovine endothelial cells." J Biol Chem. 270. 9590-9596 (1995)
[Publications] Naved, AF., et al.: "CBP-140,a novel endoplasmic reticulum resident Ca2+-binding protein with a carboxy terminal NDEL sequence showed partial homology with 70kDa heat shock protein(hsp70)." Cell Struct Funct. 20. 133-141 (1995)
[Publications] Sudou, A., et al.: "Lewis X structure increases cell substratum adhesion in L cells." J Biochem. 117. 271-275 (1995)
[Publications] Mitsiadis, T. A., et al.: "Midkine(MK),a heparin-binding growth/differentiation factor,is regulated by retinoic acid and epithelial-mesenchymal interactions in the developing mouse tooth, and affects cell proliferation and morphogenesis." J Cell Biol. 129. 267-281 (1995)
[Publications] Mitsiadis, T. A., et al.: "Expression of the heparin-binding cytokines,midkine(MK)and HB-GAM(pleiotrophin)is associated with epithelial-mesenchymal interactions during fetal development andorganogenesis." Development. 121. 37-51 (1995)

1995 Fiscal Year Annual Research Report

標的遺伝子組換えによる突然変異体の作成とその生体機能の研究

Principal Investigator

勝木 元也 東京大学, 医科学研究所, 教授 (20051732)

Research Products

[Publications] Yamaguchi, H., et al.: "Dopamine D2 receptor plays a critical role in cell proliferation and proopiomelanocortin expression in the pituitary." Genes to Cells. in press.

[Publications] Gondo, Y., et al.: "A novel positive detection system of in vivo mutations in rpsL(strA)transgenic mice." Mutation Res. in press.

[Publications] Masu, M., et al.: "Specific deficit of the ON response in visual transmission by targeted disruption of the mGluR6 gene." Cell. 80. 757-765 (1995)

[Publications] Shimano, H., et al.: "Inhibition of diet-induced atheroma formation in transgenic mice expressing apolipoprotein E in the arterial wall." J Clin Invest. 95. 469-476 (1995)

[Publications] Honda, H., et al.: "Increased tyrosine-phosphorylation of 55KDa proteins in beta-actin/Tec transgenic mice." Biochem Biophys Res Commun. 206. 287-293 (1995)

[Publications] Tamura, H., et al.: "Opaque eyes developed in transgenic mice with T-cell receptor δ gene." Invest Ophthalmol Vis Sci. 36. 467-477 (1995)

[Publications] Komano, H., et al: "Homeostatic regulation of intestinal epithelia by intraepithelial γδ T cells." Proc Natl Acad Sci USA. 92. 6147-6151 (1995)

[Publications] Gomi, H., et al.: "Mice devoid of the glial fibrillary acidic protein develop normally and are susceptible to scrapie prions." Neuron. 14. 29-41 (1995)

[Publications] Emoto, M., et al.: "Mice dificient in γδ T cells are resistant to lethal infection with Salmonella choleraesuis." Infect Immun. 63. 3736-3738 (1995)

[Publications] Nishimura, H., et al.: "The role of γδ T cells in priming macrophages to produce tumor necrosis factor-α." Eur Immunol. 25. 1465-1468 (1995)

[Publications] Nakane, H., et al.: "High incidence of UVB-or chemical carcinogen-induced skin tumorigenesis in mice lacking the xerodema pigmentosum group A(XPA)gene." Nature. in press.

[Publications] Kaneda, Y., et al.: "The recycling of synaptic vesicle membrane components. “Protein Kinesis: The Dynamics of Protein Trafficking and Stability"" Cold Spring Harbor Symp. Quant. Biol. 60(in press).

[Publications] Mann, M. J., et al.: "Genetic engineering of vein grafts resistant to atherosclerosis." Proc Natl Acad Sci USA. 92. 4502-4506 (1995)

[Publications] von der Leyen, H. E., et al.: "Gene therapy inhibiting neointimal vascular lesion: In vivo transfer of endothelial cell nitric oxide synthase gene." Proc Natl Acad Sci USA. 92. 1137-1141 (1995)

[Publications] Kaneda. Y., et al.: "Increased wxpression of DNA cointroduced with nuckear protein in adult rat liver." J Mol Med. 73. 289-297 (1995)

[Publications] Mizuki, N., et al.: "Isolation of cDNA and genomic clones of a human Ras-related GTP-binding protein gene and its choromosomal localization to the long arm of choromosome 7, 7q36." Genomics. in press.

[Publications] Tsai, HJ., et al.: "Initiation of the transgenic lacZ gene expression in medaka(Oryzias latipes)embryos." Mol Mar Biol Biotechnol. 4. 1-9 (1995)

[Publications] Naruse, T., et al.: "Cloned PLT cells recognize the forth,fifth and sixth hypervariable regions at amino acid positions 65 to 87 of DPB1 molecule." Human Immunol. 42. 123-130 (1995)

[Publications] Kajiwara, K., et al.: "Cloning and characterization of pentylenetetrazol-related cDNA, PTZ-17." Brain Res. 671. 170-174 (1995)

[Publications] Cai, J-P., et al.: "cDNA and genomic sequences for rat 8-oxo-dGTPase that prevents occurrence of spontaneous mutations due to oxidation of guanine nucleotides." Carcinogenesis. 16. 2343-2350 (1995)

[Publications] Kakuma, T., et al.: "Mouse MTH1 protein with 8-Oxo-7,8-dihydro-2′-deoxyguanosine5′-triphatase activity that prevents transversion mutation." J Biological Chem. 270. 2542-2548 (1995)

[Publications] Rancourt, D. E., et al.: "Genetic interaction between hoxb-5 and hoxb-6 is revealed by nonallelic noncomplementation." Genes Dev. 9. 108-122 (1995)

[Publications] Kuromitsu, J., et al.: "Programmed alterations of DNA methylation at CpG islands during blastformation of peripheral blood lymphocytes." DNA Res. in press.

[Publications] Plass, C., et al.: "Comparative analysis of mouse NotI Iinking clones with mouse and human genomic sequences and transcripts." DNA Res. 2. 27-35 (1995)

[Publications] Hirotsune, S., et al.: "The reeler gene encodes a protein with EGF-like motif expressed by pioneer neurons." Nature Genetics. 10. 77-83 (1995)

[Publications] Latham, K., et al.: "Stage-specific induction and regulation by genomic imprinitng of mouse U2bpaf-rs gene during preimplantation development." Developmental Biology. 168. 670-676 (1995)

[Publications] Okazaki, Y., et al.: "Direct detection and isolation of RLGS spot DNA marker tightly linked to a specific trait using RLGS spot-bombing method." Proc Natl Acad Sci USA. 92. 5610-5614 (1995)

[Publications] Ohsumi, T., et al.: "Loss of heterozygosity in chromosomes 1, 5, 7and13 in mouse hepatoma detected by systematic genome-wide scanning RLGS genetic map." Biochem Biophys Res Commun. 212. 632-639 (1995)

[Publications] Kalcheva, I., et al.: "Comparative mapping of the imprinted U2afbpL gene on mouse chromosome11 and human chromosome5." Cytogenet Cell grenet. 68. 19-24 (1995)

[Publications] Okazaki, Y, et al.: "Amultiplex procduction system of RLGS gel-A new version of restriction landmark genomic scanning method(RLGS Ver1, 8)." Electrophoresis. 16. 197-202 (1995)

[Publications] Ohsumi, T, et al.: "RLGS spot cloning method." Electrophoresis. 16. 203-209 (1995)

[Publications] Shibata, H., et al.: "Systematic screening of endogenous imprinted genes using RLGS and identification of a new imprinted gene U2af binding protein related sequence." Electrophoresis. 16. 210-217 (1995)

[Publications] Watanabe, S., et al.: "Accessibilitey to tissue-specific genes from methylantion profiles of mouse brain genomic DNA." Electrophoresis. 16. 218-226 (1995)

[Publications] Okuizumi, H., et al.: "Genetic mapping of restriction landmark senomic scanning(RLGS)loci in the mouse." Electrophoresis. 16. 233-240 (1995)

[Publications] Okuizumi, H., et al.: "A single gel analysis of 573 dominant and codominant restriction landmark genomic scanning(RLGS)loci in interspecific backcross progeny." Electrophoresis. 16. 253-260 (1995)

[Publications] Horie, K., et al.: "A replacement vector us ed to introduce subtle mutaions into mouse genes." Gene. in press.

[Publications] Kato, G. et al.: "Novel phosphory lation at a mitotic site, serine 75,in human pp60^<c-src> from unsynchronized human tumor cells having a spherical morphology." Biochem Biophys Res Commun. 216. 619-629 (1995)

[Publications] Wei, S., et al.: "Studies on the metabolism of retinol and retinol-binding protein in transthyretin-deficient mice produced by homologous recombi-nation." J Biol Chem. 270. 866-870 (1995)

[Publications] Matsuo. I., et al.: "Mouse Ot×2 functions in the formation and patteming of rostral head." Gene Dev. 9. 2646-2658 (1995)

[Publications] Miyauchi, T., et al.: "Structure of the mouse basigin gene,a unique member of the immunoglobulin superfamily." J Biochem. in press.

[Publications] Masuda, K., et al.: "Functional rescue of photoreceptos from the damaging effects of constant light by survival-promoting factors in the rat." Invest Ophtalmol Vis Sci. in press.

[Publications] Obama, H., et al.: "A transgenic mouse line with α-1,3/4 fucosyltransferase cDNA:production and characteristics." Glycoconjugate J. 12. 795-801 (1995)

[Publications] Yu, S., et al.: "cDNA cloning and sequence analysis of a novel calcium binding protein with oligoproline motif" Cell Struct Funct. 20. 263-268 (1995)

[Publications] Kojima, S., et al.: "Midkine enhances fibrinolytic activity of bovine endothelial cells." J Biol Chem. 270. 9590-9596 (1995)

[Publications] Naved, AF., et al.: "CBP-140,a novel endoplasmic reticulum resident Ca2+-binding protein with a carboxy terminal NDEL sequence showed partial homology with 70kDa heat shock protein(hsp70)." Cell Struct Funct. 20. 133-141 (1995)

[Publications] Sudou, A., et al.: "Lewis X structure increases cell substratum adhesion in L cells." J Biochem. 117. 271-275 (1995)

[Publications] Mitsiadis, T. A., et al.: "Midkine(MK),a heparin-binding growth/differentiation factor,is regulated by retinoic acid and epithelial-mesenchymal interactions in the developing mouse tooth, and affects cell proliferation and morphogenesis." J Cell Biol. 129. 267-281 (1995)

[Publications] Mitsiadis, T. A., et al.: "Expression of the heparin-binding cytokines,midkine(MK)and HB-GAM(pleiotrophin)is associated with epithelial-mesenchymal interactions during fetal development andorganogenesis." Development. 121. 37-51 (1995)

勝木元也東京大学, 医科学研究所, 教授 (20051732)