1992 Fiscal Year Annual Research Report

外来遺伝子DNA導入により新しいモデル動物の開発

Research Project

Project/Area Number	04206101
Research Institution	Kyushu University
Principal Investigator	勝木元也九州大学, 生体防御医学研究所, 教授 (20051732)
Co-Investigator(Kenkyū-buntansha)	権藤洋一東海大学, 医学部, 助手 (40225678) 森正敬熊本大学, 医学部, 教授 (40009650) 岩倉洋一郎東京大学, 医科学研究所, 助教授 (10089120) 近藤寿人名古屋大学, 理学部, 教授 (70127083) 山村研一熊本大学, 医学部, 教授 (90115197)
Keywords	トランスジェニックマウス / ジーンターゲッティング / ヒト疾患モデル / 生物機能モデル / 胚幹細胞 / 遺伝子欠損モデル / トランスサイレチン / HTLVI
Research Abstract	本研究の目的は、遺伝子導入法を駆使して新しい生物機能モデルおよびヒト疾患モデルを開発することである。また、ジーンターゲッティング法を確立し、遺伝子欠損モデルを作ることも目的の一つである。本年度は、6年間の本重点領域研究の最終年度に当たり、これまでの成果を完成する方向での研究が行われた。第1は遺伝子導入法によるトランスジェニックマウスの作成である。トランスサイレチン遺伝子のプロモーター領域を6kb以上に長くすると、肝臓など正常マウスで本来発現している臓器で特異的な発現が認められることを確認した。このマウスは通常の飼育を続けると6ヶ月以上でアミロイドの沈着が認められヒト疾患モデルとなることがわかった。しかし、清浄な施設で飼育するとアミロイドの沈着が1年後でも全く認められなかった。これは動物実験でしか明らかにならなかったものとして貴重な結果である。この他、高血圧マウス、ラット、I型糖尿病NODマウスに欠損している遺伝子導入による病態の回復、ApoE遺伝子導入による脂質代謝の亢進と動脈硬化予防との関係、HTLVIのtax遺伝子導入による関節炎の発症とリューマチとの関係などが解明され、トランスジェニック法が極めて有効なヒト疾患の解析およびモデル動物の開発法であることをあらためて示すことが出来た。第2は標的遺伝子破壊法ともいえるジーンターゲッティング法の開発と確立である。方法はすでに普及の段階に達しているが、胚幹細胞やフィーダー細胞の性質によって大きく影響される。本邦の他の分野で開発されたTT2細胞と胚由来のフィーダー細胞を用いれば、ほぼ100%に近い高率で、胚幹細胞が生殖細胞へと分化することが確認された。その結果、遺伝子トラップ法によって10以上の新しい遺伝子を取ることができた。また、N-myc.p53,NMDARなどの遺伝子が破壊されたマウスを作ることができた。

Research Products

(24 results)

All Other

All Publications (24 results)

[Publications] Sasaoka,T.: "Analysis of the human tyrosine hydroxylase promoter-chloram-phenicol acetyltransferase chimeric gene expression in transgenic mice." Mol.Brain Res.16. 274-286 (1992)
[Publications] Abeliovich,A.: "On somatic recombination in the central nervous system of transgenic mice." Science. 257. 404-408 (1992)
[Publications] Takagi,K.: "A rapid and efficient protocol of the inverted PCR using two primer pairs." Biotechniques. 13. 176-178 (1992)
[Publications] Hayashi,T.: "RNA packing signal of human immunodeficiency virus type 1." Virology. 188. 590-599 (1992)
[Publications] Iwakura,Y.: "The induction of cataracts by HIV-I in transgenic mice." AIDS. 6. 1069-1075 (1992)
[Publications] Kawahara,T.: "Ribozymes for specific inhibition of mRNA function in the nematode Caenorhabditis elegans.1" Nucleic Acids Symp Ser.27. 45-46 (1992)
[Publications] Aroian,RV.: "Splicing in Caenorhabditis elegans does not require an AG at the 3' splice acceptor site." Mol.Cell Biol.13. 626-702 (1993)
[Publications] Sasai,Y.: "Two mammalian helix-loop-helix factors structurally related to Drosophila hairy and Enhancer of Split." Genes Dev.6. 2620-2634 (1992)
[Publications] Akazawa,C.: "Molecular characterization of a rat negative regulator with a basic helix-loop-helix structure predominantly expressed in the developing nervous system." J.Biol.Chem.267. 21879-21885 (1992)
[Publications] Sawada,M.: "Stable expression of monkey cytochrome p-450IAL cDNA in Chinese hamster CHL cells and its application for detection of mutagenicity of aflatoxin B_1." Mutation Res.265. 23-29 (1992)
[Publications] Fukuta,H.: "Toxicological significance of dog liver cytochrome p-450:Examination with the enzyme expressed in Saccharomyces cerevisiae using recombinant expression plasmid." Mutation Res.269. 97-105 (1992)
[Publications] Sawai,S.: "Defects of embryonic organogenesis resulting from targeted disruption of the N-myc gene of the mouse." Development. (1993)
[Publications] Wakamatsu,Y.: "Transition of localization of the N-myc protein from nucleus to cytoplasm in differentiating neurons." Neuron. (1993)
[Publications] Gondo,Y.: "High-frequency genetic reversion mediated by a DNA duplication:The mouse pink-eyed unstable mutation." Proc.Natl.Acad.Sci.USA. 90. 297-301 (1993)
[Publications] Gardner,JM.: "The mouse pink-eyed dilution gene.Association with human Prader-Willi and Angelman Syndromes." Science. 257. 1121-1124 (1992)
[Publications] Yoshioka,K.: "Production of a unique multi-lamella structure in the nuclei of yeast expression Drosophila copia gag precursor." FEBS. 302. 5-7 (1992)
[Publications] Yoshioka,K.: "Efficient amplification of Drosophila simulans copia directed by high-level reverse transcriptase activity associated with copia virus-like particles." Gene. 120. 191-196 (1992)
[Publications] Suematsu,S.: "Generation of plasmacytomas with the chromosomal translocation t (12;15) in interleukin 6 transgenic mice." Proc.Natl.Acad.Sci.USA. 89. 232-235 (1992)
[Publications] Miyazaki,T.: "Prevention of autoimmune insulitis in nonobese diabetic mice by expression of major histocompatibility complex class II molecules." Proc.Natl.Acad.Sci.USA. 89. 9519-9523 (1992)
[Publications] Nagahata,T.: "Inhibition of intrahepatis B virus replication by antiviral drugs in a novel transgenic mouse model." Antimicro.Agents Chemother.36. 2042-2045 (1992)
[Publications] Katsuki,M.: "Chemically induced tumors in transgenic mice carrying prototype human c-Ha-ras genes.Multistage Carcinogenesis." CRC Press,Tpkyo, 249-257 (1992)
[Publications] Kimura,M.: "Manipulation of myelin formation in transgenic mice.Antisense RNA and DNA." Wiley-Liss,Inc., 109-120 (1992)
[Publications] Takiguchi,M.: "Structureand ewpression of the urea cycle enzyme genes.Frontiers and new horizons in amino acid research." Elsevier Science Publishers, 145-150 (1992)
[Publications] Takiguchi,M.: "Structure and expression of liver-type arginase gene.Guanidino compounds in biology and medicine." John Libbey&Company, 349-353 (1992)

1992 Fiscal Year Annual Research Report

外来遺伝子DNA導入により新しいモデル動物の開発

Principal Investigator

勝木 元也 九州大学, 生体防御医学研究所, 教授 (20051732)

Research Products

[Publications] Sasaoka,T.: "Analysis of the human tyrosine hydroxylase promoter-chloram-phenicol acetyltransferase chimeric gene expression in transgenic mice." Mol.Brain Res.16. 274-286 (1992)

[Publications] Abeliovich,A.: "On somatic recombination in the central nervous system of transgenic mice." Science. 257. 404-408 (1992)

[Publications] Takagi,K.: "A rapid and efficient protocol of the inverted PCR using two primer pairs." Biotechniques. 13. 176-178 (1992)

[Publications] Hayashi,T.: "RNA packing signal of human immunodeficiency virus type 1." Virology. 188. 590-599 (1992)

[Publications] Iwakura,Y.: "The induction of cataracts by HIV-I in transgenic mice." AIDS. 6. 1069-1075 (1992)

[Publications] Kawahara,T.: "Ribozymes for specific inhibition of mRNA function in the nematode Caenorhabditis elegans.1" Nucleic Acids Symp Ser.27. 45-46 (1992)

[Publications] Aroian,RV.: "Splicing in Caenorhabditis elegans does not require an AG at the 3' splice acceptor site." Mol.Cell Biol.13. 626-702 (1993)

[Publications] Sasai,Y.: "Two mammalian helix-loop-helix factors structurally related to Drosophila hairy and Enhancer of Split." Genes Dev.6. 2620-2634 (1992)

[Publications] Akazawa,C.: "Molecular characterization of a rat negative regulator with a basic helix-loop-helix structure predominantly expressed in the developing nervous system." J.Biol.Chem.267. 21879-21885 (1992)

[Publications] Sawada,M.: "Stable expression of monkey cytochrome p-450IAL cDNA in Chinese hamster CHL cells and its application for detection of mutagenicity of aflatoxin B_1." Mutation Res.265. 23-29 (1992)

[Publications] Fukuta,H.: "Toxicological significance of dog liver cytochrome p-450:Examination with the enzyme expressed in Saccharomyces cerevisiae using recombinant expression plasmid." Mutation Res.269. 97-105 (1992)

[Publications] Sawai,S.: "Defects of embryonic organogenesis resulting from targeted disruption of the N-myc gene of the mouse." Development. (1993)

[Publications] Wakamatsu,Y.: "Transition of localization of the N-myc protein from nucleus to cytoplasm in differentiating neurons." Neuron. (1993)

[Publications] Gondo,Y.: "High-frequency genetic reversion mediated by a DNA duplication:The mouse pink-eyed unstable mutation." Proc.Natl.Acad.Sci.USA. 90. 297-301 (1993)

[Publications] Gardner,JM.: "The mouse pink-eyed dilution gene.Association with human Prader-Willi and Angelman Syndromes." Science. 257. 1121-1124 (1992)

[Publications] Yoshioka,K.: "Production of a unique multi-lamella structure in the nuclei of yeast expression Drosophila copia gag precursor." FEBS. 302. 5-7 (1992)

[Publications] Yoshioka,K.: "Efficient amplification of Drosophila simulans copia directed by high-level reverse transcriptase activity associated with copia virus-like particles." Gene. 120. 191-196 (1992)

[Publications] Suematsu,S.: "Generation of plasmacytomas with the chromosomal translocation t (12;15) in interleukin 6 transgenic mice." Proc.Natl.Acad.Sci.USA. 89. 232-235 (1992)

[Publications] Miyazaki,T.: "Prevention of autoimmune insulitis in nonobese diabetic mice by expression of major histocompatibility complex class II molecules." Proc.Natl.Acad.Sci.USA. 89. 9519-9523 (1992)

[Publications] Nagahata,T.: "Inhibition of intrahepatis B virus replication by antiviral drugs in a novel transgenic mouse model." Antimicro.Agents Chemother.36. 2042-2045 (1992)

[Publications] Katsuki,M.: "Chemically induced tumors in transgenic mice carrying prototype human c-Ha-ras genes.Multistage Carcinogenesis." CRC Press,Tpkyo, 249-257 (1992)

[Publications] Kimura,M.: "Manipulation of myelin formation in transgenic mice.Antisense RNA and DNA." Wiley-Liss,Inc., 109-120 (1992)

[Publications] Takiguchi,M.: "Structureand ewpression of the urea cycle enzyme genes.Frontiers and new horizons in amino acid research." Elsevier Science Publishers, 145-150 (1992)

[Publications] Takiguchi,M.: "Structure and expression of liver-type arginase gene.Guanidino compounds in biology and medicine." John Libbey&Company, 349-353 (1992)

勝木元也九州大学, 生体防御医学研究所, 教授 (20051732)