1993 年度実績報告書

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

研究課題

研究課題/領域番号	05275102
研究機関	九州大学
研究代表者	勝木元也九州大学, 生体防御医学研究所, 教授 (20051732)
研究分担者	前田秀一郎山梨医科大学, 教授 (10117244) 木村穣東海大学, 医学部, 助教授 (10146706) 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, ウイルス研究所, 助教授 (60252524) 相沢慎一理化学研究所ライフサイエンス筑波研究センター, 副主任研究員 (60073011)
キーワード	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 / セロトニン受容体 / p53
研究概要	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,個体レベルでの解析が必須であると考えられる中枢神経系,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,マウスの遺伝子を制限酵素で分解した後,二次元展開して,新しい遺伝子を見出した林崎班員の成果は即刻勝木のもとでジーンターゲッティングがなされるなど領域設定が大いに意義あるものとなっている。本年度において破壊された遺伝子を列挙すれば次の如くである。グルタミン酸受容体(NMDAタイプ5種およびメタボトロピック6種):ホモ型突然変異体のうちNMDA受容体1(NR1)破壊マウスは生後1日目にミルクが飲めずに死亡する。セロトニン受容体:現在キメラマウス。p53遺伝子をbeta-ガラクトシダーゼ遺伝子で置換したマウス:p53の発現を受精から成体まで染色によって解析でき,その特徴的な発現パターンを研究中である。GFAP,XPAC,Fyn,CSK,テネシンなどの遺伝子が破壊された。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るものは少なく,それぞれの遺伝子産物が多くの生体機能に関っていることが示された。本年度は本領域の初年度であるに拘らず,多くの大きな成果が得られたと評価できる。

研究成果

(32件)

すべてその他

すべて文献書誌 (32件)

[文献書誌] Akiyama,M. Katsuki,M.,et al.: "Lymphocyte infiltration of the skin in transgenic mice carrying the human interleukin-2 gene." Arch Dermatol Res. 285. 379-384 (1993)
[文献書誌] Takaori,K. Katsuki,M.,et al.: "Biochemical characteristics of human renin expressed in transgenic mice." Clinical Science. 84. 21-29 (1993)
[文献書誌] Hayashizaki,Y. Katsuki,M.,et al.: "Identification of an imprinted U2af binding protein related sequence on mouse chromosome 11 using RLGS method." Nature Genetics. 6. 33-40 (1993)
[文献書誌] Miyado,K. Katsuki,M.,et al.: "Isolation of a yeast cDNA clone that encodes a novel transmembrane protein having a c-terminal highly basic region." Biochem Biophys Res Commun. in press. (1994)
[文献書誌] Imoto,S. Aizawa,S.,et al: "Regulation of class II MHC gene expression by the inducible antisense RNA in transgenic mice." Anal Biochem. 210. 231-234 (1993)
[文献書誌] Furuta,Y. Aizawa,S.,et al: "Degeneration of skeletal and cardiac muscles in c-myb transgenic mice." Transgenic Res. 2. 199-207 (1993)
[文献書誌] Yagi,T. Aizawa,S.et al.: "A novel negative selection for homologous recombinants using diphtheria toxin A a fragment gene." Anal Biochem. 214. 77-86 (1993)
[文献書誌] Yagi,T. Aizawa,S.et al.: "A novel ES cell line,TT2,with high germline-differentiatin potency." Anal Biochem. 214. 70-76 (1993)
[文献書誌] Yagi,T. Aizawa,S.,et al: "A novel negative selection for homologous recombinants using diphteria toxin A fragment gene." Anal Biochem. 214. 379-384 (1993)
[文献書誌] Yagi,T. Aizawa,S.: "A novel ES cell line,TT2,with high germline-differentiating potency." Anal Biochem. 214. 70-76 (1993)
[文献書誌] Yagi,T. Aizawa,S. et al.: "Regional localization of Fyn in adult brain;studies with mice in which fyn gene was replaced by lacZ." Oncogene. 8. 3343-3351 (1993)
[文献書誌] Tsukada,T. Aizawa,S. et al.: "Enhanced proliferative potential in culture of cells from p53-deficient mice." Oncogene. 8. 3313-3322 (1993)
[文献書誌] Yagi,T. Aizawa,S.et al.: "A role for Fin tyrosine kinase in the suckling behavior of neonatal mice." Nature. 366. 742-745 (1993)
[文献書誌] Itohara,S.et al.: "T-cell receptor delta gene mutant mice:Independent generation of gamma delta T cells and programmed rearrangements of gamma delta TCR genes." Cell. 72. 337-348 (1993)
[文献書誌] Gomi.H. Itohara.S.et al.: "Prion protein(PrP)is not involved in the pathogenesis of spongiform encephalopathy in zitter rats." Neuroscience Letters. (in press). (1994)
[文献書誌] Isaka,Y. Kaneda,Y.et al.: "Glomerulosclerosis induced by in vivo transfection with TGF beta-or PDGF gene into rat kidney." J Clin Invest. (in press). (1994)
[文献書誌] Morishita,R. Kaneda,Y.: "In vivo gene transfer into intact blood vessels by HVJ method." Circulation. (in press). (1994)
[文献書誌] Tomita,N. Kaneda,Y.et al.: "A novel gene transfer technique mediated by HVJ (Sendai Virus),nuclear protein and liposomes." Cancer Detection and Prevention. in press. (1994)
[文献書誌] Morishita,R. Kaneda,Y.et al.: "Intimal hyperplasia after vascular injury is inhibited by antisence CDK2 kinase oligonucleotide." J Clin Invest. in press. (1994)
[文献書誌] Kaneda,Y. et al.: "The analysis of 40 kDa nuclear protein,p40,in interphase cells and mitotic cells." J Cell Sci. 106. 741-748 (1993)
[文献書誌] Morishita,R. Kaneda,Y.et al.: "Single intraluminal delivery of antisense cdc2 kinase and proliferating-cell nuclear antigen oligonucleotides results in chronic inhibition of neointimal hyperplasia." Proc Natl.Acad Sci USA. 90. 8474-8478 (1993)
[文献書誌] Uchiyama,A. Kaneda,Y. et al.: "Transfection of interleukin-2 gene into human melanoma cells augments cellular immune response." Cancer Res.53. 949-952 (1993)
[文献書誌] Tomita,N. Kaneda,Y.et al.: "Hypertensive rats produced by in vivo introduction of the human renin gene." Circulation Res. 73. 898-905 (1993)
[文献書誌] Morishita,R. Kaneda,Y.et al.: "Novel in vitro gene transfer method for study of local modulators in vascular smooth muscle cells." Hypertension. 21. 894-899 (1993)
[文献書誌] Morishita,R. Kaneda,Y.et al.: "Novel and effective gene transfer technique for study of vascular renin angiotensin system." J Clin Invest. 91. 2580-2585 (1993)
[文献書誌] Takagi,S. Kimura,M. et al.: "Direct sequencing of PCR products using unlabelled primers." BioTechniques. 14. 218-221 (1993)
[文献書誌] Hayashizaki,Y. Katsuki,M. et al.: "Restriction landmark genomic scanning and its various application." Electrophoresis. 14. 251-258 (1993)
[文献書誌] Hirotsune,S. Hayashizaki,Y.et al.: "Molecular cloning of polymorphoic markers on RLGS gel using the spot target cloning method." Biochem Biophys Res Commun. 194. 1406-1412
[文献書誌] Nagai,H.Hayashizaki,Y.et al: "Genomic analysis of human hepatocellular carcinomas using restriction landmark genomic scanning." Cancer Detect. Prevention. 17. 399-404 (1993)
[文献書誌] Kawai,J.Hayashizaki,Y.et al.: "Methylation profile of genomic DNA mouse developmental brain detected by restriction landmark genomic scanning(RLGS) method." Nucl Acid Res. 21. 5604-5608 (1993)
[文献書誌] 勝木元也: "神経疾患の分子医学「高次神経機能を遺伝子から視る」" 羊土社, 148 (1993)
[文献書誌] 前田秀一郎: "神経疾患の分子医学「優性遺伝病、家族性アミロイドポリニューロパチーのモデルマウスの作出」" 羊土社, 148 (1993)

1993 年度 実績報告書

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

研究代表者

勝木 元也 九州大学, 生体防御医学研究所, 教授 (20051732)

研究成果

[文献書誌] Akiyama,M. Katsuki,M.,et al.: "Lymphocyte infiltration of the skin in transgenic mice carrying the human interleukin-2 gene." Arch Dermatol Res. 285. 379-384 (1993)

[文献書誌] Takaori,K. Katsuki,M.,et al.: "Biochemical characteristics of human renin expressed in transgenic mice." Clinical Science. 84. 21-29 (1993)

[文献書誌] Hayashizaki,Y. Katsuki,M.,et al.: "Identification of an imprinted U2af binding protein related sequence on mouse chromosome 11 using RLGS method." Nature Genetics. 6. 33-40 (1993)

[文献書誌] Miyado,K. Katsuki,M.,et al.: "Isolation of a yeast cDNA clone that encodes a novel transmembrane protein having a c-terminal highly basic region." Biochem Biophys Res Commun. in press. (1994)

[文献書誌] Imoto,S. Aizawa,S.,et al: "Regulation of class II MHC gene expression by the inducible antisense RNA in transgenic mice." Anal Biochem. 210. 231-234 (1993)

[文献書誌] Furuta,Y. Aizawa,S.,et al: "Degeneration of skeletal and cardiac muscles in c-myb transgenic mice." Transgenic Res. 2. 199-207 (1993)

[文献書誌] Yagi,T. Aizawa,S.et al.: "A novel negative selection for homologous recombinants using diphtheria toxin A a fragment gene." Anal Biochem. 214. 77-86 (1993)

[文献書誌] Yagi,T. Aizawa,S.et al.: "A novel ES cell line,TT2,with high germline-differentiatin potency." Anal Biochem. 214. 70-76 (1993)

[文献書誌] Yagi,T. Aizawa,S.,et al: "A novel negative selection for homologous recombinants using diphteria toxin A fragment gene." Anal Biochem. 214. 379-384 (1993)

[文献書誌] Yagi,T. Aizawa,S.: "A novel ES cell line,TT2,with high germline-differentiating potency." Anal Biochem. 214. 70-76 (1993)

[文献書誌] Yagi,T. Aizawa,S. et al.: "Regional localization of Fyn in adult brain;studies with mice in which fyn gene was replaced by lacZ." Oncogene. 8. 3343-3351 (1993)

[文献書誌] Tsukada,T. Aizawa,S. et al.: "Enhanced proliferative potential in culture of cells from p53-deficient mice." Oncogene. 8. 3313-3322 (1993)

[文献書誌] Yagi,T. Aizawa,S.et al.: "A role for Fin tyrosine kinase in the suckling behavior of neonatal mice." Nature. 366. 742-745 (1993)

[文献書誌] Itohara,S.et al.: "T-cell receptor delta gene mutant mice:Independent generation of gamma delta T cells and programmed rearrangements of gamma delta TCR genes." Cell. 72. 337-348 (1993)

[文献書誌] Gomi.H. Itohara.S.et al.: "Prion protein(PrP)is not involved in the pathogenesis of spongiform encephalopathy in zitter rats." Neuroscience Letters. (in press). (1994)

[文献書誌] Isaka,Y. Kaneda,Y.et al.: "Glomerulosclerosis induced by in vivo transfection with TGF beta-or PDGF gene into rat kidney." J Clin Invest. (in press). (1994)

[文献書誌] Morishita,R. Kaneda,Y.: "In vivo gene transfer into intact blood vessels by HVJ method." Circulation. (in press). (1994)

[文献書誌] Tomita,N. Kaneda,Y.et al.: "A novel gene transfer technique mediated by HVJ (Sendai Virus),nuclear protein and liposomes." Cancer Detection and Prevention. in press. (1994)

[文献書誌] Morishita,R. Kaneda,Y.et al.: "Intimal hyperplasia after vascular injury is inhibited by antisence CDK2 kinase oligonucleotide." J Clin Invest. in press. (1994)

[文献書誌] Kaneda,Y. et al.: "The analysis of 40 kDa nuclear protein,p40,in interphase cells and mitotic cells." J Cell Sci. 106. 741-748 (1993)

[文献書誌] Morishita,R. Kaneda,Y.et al.: "Single intraluminal delivery of antisense cdc2 kinase and proliferating-cell nuclear antigen oligonucleotides results in chronic inhibition of neointimal hyperplasia." Proc Natl.Acad Sci USA. 90. 8474-8478 (1993)

[文献書誌] Uchiyama,A. Kaneda,Y. et al.: "Transfection of interleukin-2 gene into human melanoma cells augments cellular immune response." Cancer Res.53. 949-952 (1993)

[文献書誌] Tomita,N. Kaneda,Y.et al.: "Hypertensive rats produced by in vivo introduction of the human renin gene." Circulation Res. 73. 898-905 (1993)

[文献書誌] Morishita,R. Kaneda,Y.et al.: "Novel in vitro gene transfer method for study of local modulators in vascular smooth muscle cells." Hypertension. 21. 894-899 (1993)

[文献書誌] Morishita,R. Kaneda,Y.et al.: "Novel and effective gene transfer technique for study of vascular renin angiotensin system." J Clin Invest. 91. 2580-2585 (1993)

[文献書誌] Takagi,S. Kimura,M. et al.: "Direct sequencing of PCR products using unlabelled primers." BioTechniques. 14. 218-221 (1993)

[文献書誌] Hayashizaki,Y. Katsuki,M. et al.: "Restriction landmark genomic scanning and its various application." Electrophoresis. 14. 251-258 (1993)

[文献書誌] Hirotsune,S. Hayashizaki,Y.et al.: "Molecular cloning of polymorphoic markers on RLGS gel using the spot target cloning method." Biochem Biophys Res Commun. 194. 1406-1412

[文献書誌] Nagai,H.Hayashizaki,Y.et al: "Genomic analysis of human hepatocellular carcinomas using restriction landmark genomic scanning." Cancer Detect. Prevention. 17. 399-404 (1993)

[文献書誌] Kawai,J.Hayashizaki,Y.et al.: "Methylation profile of genomic DNA mouse developmental brain detected by restriction landmark genomic scanning(RLGS) method." Nucl Acid Res. 21. 5604-5608 (1993)

[文献書誌] 勝木元也: "神経疾患の分子医学 「高次神経機能を遺伝子から視る」" 羊土社, 148 (1993)

[文献書誌] 前田秀一郎: "神経疾患の分子医学 「優性遺伝病、家族性アミロイドポリニューロパチーのモデルマウスの作出」" 羊土社, 148 (1993)

1993 年度実績報告書

勝木元也九州大学, 生体防御医学研究所, 教授 (20051732)

[文献書誌] 勝木元也: "神経疾患の分子医学「高次神経機能を遺伝子から視る」" 羊土社, 148 (1993)

[文献書誌] 前田秀一郎: "神経疾患の分子医学「優性遺伝病、家族性アミロイドポリニューロパチーのモデルマウスの作出」" 羊土社, 148 (1993)