1993 年度実績報告書

ヒト・ゲノム解析研究

研究課題

研究課題/領域番号	05NP0401
研究機関	大阪大学
研究代表者	松原謙一大阪大学, 細胞生体工学センター, 教授 (20037394)
研究分担者	大木操国立がんセンター研究所, 部長 (00158792) 中村祐輔 (財)癌研究会癌研究所, 部長 (70217909) 吉田光昭東京大学, 医科学研究所, 教授 (80012607) 吉川寛奈良先端科学技術大学院大学, 教授 (70019876) 榊佳之東京大学, 医科学研究所, 教授 (10112327)
キーワード	ヒト・ゲノム / ゲノム構造解析 / ゲノム機能解析 / cDNA / DNA解析技術 / モデル生物 / 生物情報科学 / 塩基配列決定
研究概要	生命の設計図であるゲノムの構造と機能を解析研究しそこに包含される遺伝情報を理解する。この目的で、総括班ならびに3つの研究班を組織して研究の推進を図り次の成果を得た。 1.総括班。全体としての活動を円滑に進めるため各種会合の企画立案調整、機器の計画的購入と配分、情報交流、調査などを積極的に進めた。また、社会との接点の問題研究の展開を図った。さらに、評価委員会を設置して全班員の業績評価を行い、その結果に基き平成5年度に向けて班員の入れ替えを行い、先鋭的なゲノム研究の活性化を図った。 2.ヒト・ゲノム構造解析班。ヒト染色体21番につき物理地図を作製したほか、国際協力によるYACコンティグマップ作りに貢献した。また、染色体3,6,8,11,X,Yにつき高密度のコスミドマップ作製を進め、ゲノムデータベース作成において国際貢献を果たした。これらの研究作業を通して大量の資料を扱いつつそれらを処理、分析し、データ化して整理する方法論の開発と実行が軌道に乗った。また、これらの成果を活用して、がん抑制遺伝子や先天性発育遅滞に関する遺伝子の単離、または位置づけに成功した。これと並んで、特定領域として免疫グロブリン遺伝子H鎖V領域とHLA遺伝子群を選び、詳細なマップ作りと遺伝子の同定を実行した。これは、来るべきシーケンシング時代のモデルとなる先導的研究である。 3.cDNA解析班。ヒトの各種臓器や細胞の中で発現している全遺伝子をカタログ化し、その発現頻度を解析し情報化する「3'特異的cDNAライブラリー解析」を進め、約25000クローンの解析を終えた。これは、世界に先駆けてゲノム研究を構造解析と機能解析の両面から行おうという新しい方向性を与えるものであり、追随する仕事が多く現われた。良質なcDNAライブラリーの作成やその迅速な分類、カタログ化の手法がほぼ完成した。また、大容量のデータ整理などにおける新しい手法が種々開発されると共に、5'特異的cDNAライブラリーの作成や、cDNAの全長解析、cDNAのゲノムへのマッピングなどを可能にするいろいろな技術開発が行われ、一部でデータ生産活動が始まった。 4.DNA解析技術開発班。DNAの取り扱い技術とマーカーDNAの染色体上へのマッピングに関わる技術開発を進めた。特に、色素プローブを用いる染色体上への位置決め技術に大幅な進歩を見た。また、ゲノムスキャンニング法の確立と応用、T3ファージによるDNA欠失セットの作成技術、超高分子DNAの電場による分離技術の確立など、本班の業績が次々と出された。これと並んで、枯草菌、分裂酵母、線虫をモデル生物として選び、国際協力によってゲノムデータベースを作りつつ技術開発とデータベース間の相互連携をつくるシステム作りが長足の進歩を遂げた。全体として、近々に具体的にシーケンスデータを大量に生産するゲノム研究第二期が近づいたと思われる。

研究成果
(40件)

すべてその他

すべて文献書誌 (40件)

[文献書誌] Virgilio,L.(Isobe,M.): "Chromosome walking on the tcl-llocus involved in T-cell neoplasia" Proc.Natl.Acad.Sci.90. 9275-9279 (1993)
[文献書誌] Sugimura,K.(Inoko,H.): "Analysis of genes within the HLA region affecting susceptibility to Ulcerative Colitis" Hum.Immunol.36. 112-118 (1993)
[文献書誌] Akao,Y.(Imai,T): "Long-range mapping of the 11q23 region involved in chromosome aberrations in human tumors by pulsed-field gel electrophoresis with a yeast artificial chromosome" Genes,Chromosomes & Cancer. 8. 167-171 (1993)
[文献書誌] Ichikawa,H.(Ohki,M.): "A NotI restriction map of the entire long arm of human chromosome 21" Nature Genet.4. 361-366 (1993)
[文献書誌] Takeda,O.(Kaneko,Y.): "There may be two tumor suppressor genes on chromosome arm 1p closely associated with biologically distinct subtypes of neuroblastoma" Genes Chrom.Cancer,in press.
[文献書誌] Harada,H.(Kimura,A.): "A missense mutation of cardiac beta-myosin heavy chain gene linked to familial hypertrophic cardiomyopathy in affected Japanese families" Biochem.Biophys.Res.Commun.194. 791-798 (1993)
[文献書誌] Nagafuchi,S.(Kondo,I.): "Dentatorubral-pallidoluysian atropy expansion of an unstable CAG trinucleotide on chromosome 12p" Nature Genet.6. 14-18 (1994)
[文献書誌] Hattori,M.(Sakaki,Y.): "Sequence-tagged NotI sites of human chromosome 21 : Sequence analysis and mapping" Genomics. 17. 39-44 (1993)
[文献書誌] Hayasaka,K.(Shimizu,N.): "Charcot-Marie-Tooth neuropathy type 1B is associated with mutations of the myelin P gene" Nature Genet.5. 31-34 (1993)
[文献書誌] Koide,R.(Tsuji,S.): "Unstable expansion of CAG repeat in hereditary dentatorubral-pallidolusian atrophy(DRPLA)" Nature Genet.6(1). 9-13 (1994)
[文献書誌] Fukutani,K.(Nakagome,Y.): "Detection of the testis determining factor in as XX man" The Journal of Urology. 149. 126-128 (1993)
[文献書誌] Toda,T.(Nakamura,Y): "Localization of a gene responsible for Fukuyama type congenital muscular dystrophy (FCMD)to chromosome 9q31-33 by linkage analysis" Nature Genetics. 5. 283-286 (1993)
[文献書誌] Jinno,Y.(Niikawa,N.): "Imprinting polymorphism of the human WT1 gene" Nature Genet.,in press.
[文献書誌] Sekiya,T.(Hayashi,K.): "Detection of DNA aberrations in human cancers by single-strand conformation polymorphism analysis of the polymerase chain reaction" Int.J.Pancreatology. 13. 69 (1993)
[文献書誌] Yoshikawa,H.(Fujiyama,A.): "Two-dementional gel electrophoretograms of human chromosome specific DNA fragments" Biochem.Biophys.Res.Commun.196. 1566-1572 (1993)
[文献書誌] Masumoto,H.: "Properties of CENP-B and its target sequence in satellite DNA" NAT ASI Series : Chromosome Segregataion and Aneuploidy. 72. 31-43 (1993)
[文献書誌] Haire,R.(Matsuda,F.): "Diversification,not utilization,of the immunoglobulin V_H gene repertoire is restricted in DiGeorge syndrome" J.Exp.Med.178. 825-834 (1993)
[文献書誌] Nagkura,J.(Miki,T): "Close linkage of the gene for Werner's syndrome to ANKI and D8S87 on the short arm of chromosome 8" Gerontol. 39. 11-15 (1993)
[文献書誌] Sasaki,Y.(Ayusawa,D.,Oishi,M.): "Construction of normalized cDNA library by introduction of a semi-solid mRNA-cDNA hybridization system" Nucl.Acid.Res.,in press.
[文献書誌] Matsubara,K.and Okubo,K.: "Identification of new genes by systematic analysis of cDNA and data base construction approach" Current Opinion in Biotechnology. 4. 672-677 (1993)
[文献書誌] Sekine,S.(Kato,S.): "Synthesis of full-length cDNA using DNA-capped mRNA" Nucl.Acids.Res.Symp.Ser. 29. 143-144 (1993)
[文献書誌] Tabara,H.(Kohara,Y.): "In situ hybridization on embryos -- Analysis of candidate clones for differentially expressed genes" The Worm Breeder's Gazette. 13. 22 (1994)
[文献書誌] Maruyama,K.and Sugano,S.: "Oligo-capping: a simple method to replace the cap structure of eukaryotic on RNA with oligoribonucleotide" Gene,in press.
[文献書誌] Seo,Y.(Nakayama,T.): "Immunoaffinity purification and characterization of a liver-specific antigen" Jpn.J.Legal Med.47. 1-5 (1993)
[文献書誌] Nomura,N.: "Prediction of the coding sequences of unidentified human genes" DNA Research,in press. 1. (1994)
[文献書誌] Inazawa,J.(Abe,T.): "High-resolution ordering of DNA markers by multi color fluorescent in situ hybridization of prophase chromosomes" Cytogenet.Cell Genet. 65. 130-135 (1994)
[文献書誌] Sugaya,K.(Ikemura,T.): Three genes(receptor gene for AGE,homeobox-containing gene PBX,and int-3 related Notch-family gene)found in the HLA class III region near the border to the class II on 6p21.3. Cytogenet.Cell Genet,in press.
[文献書誌] 河野享子 (今本文男): "YACクローンの安定化" 蛋白質核酸酵素「ゲノム解析研究」. 38. 607-612 (1993)
[文献書誌] Yokota,H.(Oishi,M.): "A differential cloning procedure of complex genomic DNA fragment" Analytical Biochem,in press.
[文献書誌] Ogasawara,N.: "Systematic sequencing of the Bacillus subtilis chromosome containing the replication origin" DNA research. 1. 1-14 (1994)
[文献書誌] Okumura,K.: "Detection by fluorescence in situ hybridization of the erythropoietin gene introduced into animal cells" Animal Cell Technology : Basic & Applied Aspects. 5. 107-113 (1993)
[文献書誌] 陶山明: "オリゴヌクレオチドを用いたシーケンシング法" 蛋白質核酸酵素. 38. 647-657 (1993)
[文献書誌] Takahashi,E.: "A high-resolution cytogenetic map of human chromosome 12 : localization of 195 new cosmid markers by direct R-banding fluorescence in situ" Hum.Genet.92. 405-409 (1993)
[文献書誌] Masubuchi,Y.(Doi,M.): "Periodic behavior of DNA molecules during steady field gel electrophoresis" Macromolecules. 26. 5269-5270 (1993)
[文献書誌] Mizukami,T.(Niwa,O.): "A 13kb resolution cosmid map of the 14Mb fission yeast genome by nonrandom sequence-tagged site mapping" Cell. 73. 121-132 (1993)
[文献書誌] Baba,Y.: "Separation of DNA fragments by high-performance liquid chromatograpy and capillary electrophoresis" J.Liq.Chromatogr.16. 955-965 (1993)
[文献書誌] Hayashizaki,Y.: "Restriction Landmark Genomic Scanning and its various application,"Changing Direction in DNA"" Electrophoresis. 14. 251 (1993)
[文献書誌] Hirai,M.: "A method gor simultaneous detection of flourescent G-bands and in situ hybridization signals" Cytogenet.Cell Genetics,in press.
[文献書誌] 加藤尚子 (藤澤久雄): "T3ファージを用いた巨大DNAのクローニングと構造解析" 蛋白質核酸酵素「ゲノム解析研究」. 38. 618-625 (1993)
[文献書誌] Kagiyama,N.(Yoshida,M.C): "A novel fluorescent method for in situ hybridization" Acta Histochem.Cytochem.26. 441-445 (1993)

1993 年度 実績報告書

ヒト・ゲノム解析研究

研究代表者

松原 謙一 大阪大学, 細胞生体工学センター, 教授 (20037394)

研究成果

[文献書誌] Virgilio,L.(Isobe,M.): "Chromosome walking on the tcl-llocus involved in T-cell neoplasia" Proc.Natl.Acad.Sci.90. 9275-9279 (1993)

[文献書誌] Sugimura,K.(Inoko,H.): "Analysis of genes within the HLA region affecting susceptibility to Ulcerative Colitis" Hum.Immunol.36. 112-118 (1993)

[文献書誌] Akao,Y.(Imai,T): "Long-range mapping of the 11q23 region involved in chromosome aberrations in human tumors by pulsed-field gel electrophoresis with a yeast artificial chromosome" Genes,Chromosomes & Cancer. 8. 167-171 (1993)

[文献書誌] Ichikawa,H.(Ohki,M.): "A NotI restriction map of the entire long arm of human chromosome 21" Nature Genet.4. 361-366 (1993)

[文献書誌] Takeda,O.(Kaneko,Y.): "There may be two tumor suppressor genes on chromosome arm 1p closely associated with biologically distinct subtypes of neuroblastoma" Genes Chrom.Cancer,in press.

[文献書誌] Harada,H.(Kimura,A.): "A missense mutation of cardiac beta-myosin heavy chain gene linked to familial hypertrophic cardiomyopathy in affected Japanese families" Biochem.Biophys.Res.Commun.194. 791-798 (1993)

[文献書誌] Nagafuchi,S.(Kondo,I.): "Dentatorubral-pallidoluysian atropy expansion of an unstable CAG trinucleotide on chromosome 12p" Nature Genet.6. 14-18 (1994)

[文献書誌] Hattori,M.(Sakaki,Y.): "Sequence-tagged NotI sites of human chromosome 21 : Sequence analysis and mapping" Genomics. 17. 39-44 (1993)

[文献書誌] Hayasaka,K.(Shimizu,N.): "Charcot-Marie-Tooth neuropathy type 1B is associated with mutations of the myelin P gene" Nature Genet.5. 31-34 (1993)

[文献書誌] Koide,R.(Tsuji,S.): "Unstable expansion of CAG repeat in hereditary dentatorubral-pallidolusian atrophy(DRPLA)" Nature Genet.6(1). 9-13 (1994)

[文献書誌] Fukutani,K.(Nakagome,Y.): "Detection of the testis determining factor in as XX man" The Journal of Urology. 149. 126-128 (1993)

[文献書誌] Toda,T.(Nakamura,Y): "Localization of a gene responsible for Fukuyama type congenital muscular dystrophy (FCMD)to chromosome 9q31-33 by linkage analysis" Nature Genetics. 5. 283-286 (1993)

[文献書誌] Jinno,Y.(Niikawa,N.): "Imprinting polymorphism of the human WT1 gene" Nature Genet.,in press.

[文献書誌] Sekiya,T.(Hayashi,K.): "Detection of DNA aberrations in human cancers by single-strand conformation polymorphism analysis of the polymerase chain reaction" Int.J.Pancreatology. 13. 69 (1993)

[文献書誌] Yoshikawa,H.(Fujiyama,A.): "Two-dementional gel electrophoretograms of human chromosome specific DNA fragments" Biochem.Biophys.Res.Commun.196. 1566-1572 (1993)

[文献書誌] Masumoto,H.: "Properties of CENP-B and its target sequence in satellite DNA" NAT ASI Series : Chromosome Segregataion and Aneuploidy. 72. 31-43 (1993)

[文献書誌] Haire,R.(Matsuda,F.): "Diversification,not utilization,of the immunoglobulin V_H gene repertoire is restricted in DiGeorge syndrome" J.Exp.Med.178. 825-834 (1993)

[文献書誌] Nagkura,J.(Miki,T): "Close linkage of the gene for Werner's syndrome to ANKI and D8S87 on the short arm of chromosome 8" Gerontol. 39. 11-15 (1993)

[文献書誌] Sasaki,Y.(Ayusawa,D.,Oishi,M.): "Construction of normalized cDNA library by introduction of a semi-solid mRNA-cDNA hybridization system" Nucl.Acid.Res.,in press.

[文献書誌] Matsubara,K.and Okubo,K.: "Identification of new genes by systematic analysis of cDNA and data base construction approach" Current Opinion in Biotechnology. 4. 672-677 (1993)

[文献書誌] Sekine,S.(Kato,S.): "Synthesis of full-length cDNA using DNA-capped mRNA" Nucl.Acids.Res.Symp.Ser. 29. 143-144 (1993)

[文献書誌] Tabara,H.(Kohara,Y.): "In situ hybridization on embryos -- Analysis of candidate clones for differentially expressed genes" The Worm Breeder's Gazette. 13. 22 (1994)

[文献書誌] Maruyama,K.and Sugano,S.: "Oligo-capping: a simple method to replace the cap structure of eukaryotic on RNA with oligoribonucleotide" Gene,in press.

[文献書誌] Seo,Y.(Nakayama,T.): "Immunoaffinity purification and characterization of a liver-specific antigen" Jpn.J.Legal Med.47. 1-5 (1993)

[文献書誌] Nomura,N.: "Prediction of the coding sequences of unidentified human genes" DNA Research,in press. 1. (1994)

[文献書誌] Inazawa,J.(Abe,T.): "High-resolution ordering of DNA markers by multi color fluorescent in situ hybridization of prophase chromosomes" Cytogenet.Cell Genet. 65. 130-135 (1994)

[文献書誌] Sugaya,K.(Ikemura,T.): Three genes(receptor gene for AGE,homeobox-containing gene PBX,and int-3 related Notch-family gene)found in the HLA class III region near the border to the class II on 6p21.3. Cytogenet.Cell Genet,in press.

[文献書誌] 河野 享子 (今本 文男): "YACクローンの安定化" 蛋白質核酸酵素「ゲノム解析研究」. 38. 607-612 (1993)

[文献書誌] Yokota,H.(Oishi,M.): "A differential cloning procedure of complex genomic DNA fragment" Analytical Biochem,in press.

[文献書誌] Ogasawara,N.: "Systematic sequencing of the Bacillus subtilis chromosome containing the replication origin" DNA research. 1. 1-14 (1994)

[文献書誌] Okumura,K.: "Detection by fluorescence in situ hybridization of the erythropoietin gene introduced into animal cells" Animal Cell Technology : Basic & Applied Aspects. 5. 107-113 (1993)

[文献書誌] 陶山 明: "オリゴヌクレオチドを用いたシーケンシング法" 蛋白質核酸酵素. 38. 647-657 (1993)

[文献書誌] Takahashi,E.: "A high-resolution cytogenetic map of human chromosome 12 : localization of 195 new cosmid markers by direct R-banding fluorescence in situ" Hum.Genet.92. 405-409 (1993)

[文献書誌] Masubuchi,Y.(Doi,M.): "Periodic behavior of DNA molecules during steady field gel electrophoresis" Macromolecules. 26. 5269-5270 (1993)

[文献書誌] Mizukami,T.(Niwa,O.): "A 13kb resolution cosmid map of the 14Mb fission yeast genome by nonrandom sequence-tagged site mapping" Cell. 73. 121-132 (1993)

[文献書誌] Baba,Y.: "Separation of DNA fragments by high-performance liquid chromatograpy and capillary electrophoresis" J.Liq.Chromatogr.16. 955-965 (1993)

[文献書誌] Hayashizaki,Y.: "Restriction Landmark Genomic Scanning and its various application,"Changing Direction in DNA"" Electrophoresis. 14. 251 (1993)

[文献書誌] Hirai,M.: "A method gor simultaneous detection of flourescent G-bands and in situ hybridization signals" Cytogenet.Cell Genetics,in press.

[文献書誌] 加藤 尚子 (藤澤 久雄): "T3ファージを用いた巨大DNAのクローニングと構造解析" 蛋白質核酸酵素「ゲノム解析研究」. 38. 618-625 (1993)

[文献書誌] Kagiyama,N.(Yoshida,M.C): "A novel fluorescent method for in situ hybridization" Acta Histochem.Cytochem.26. 441-445 (1993)

1993 年度実績報告書

松原謙一大阪大学, 細胞生体工学センター, 教授 (20037394)

[文献書誌] 河野享子 (今本文男): "YACクローンの安定化" 蛋白質核酸酵素「ゲノム解析研究」. 38. 607-612 (1993)

[文献書誌] 陶山明: "オリゴヌクレオチドを用いたシーケンシング法" 蛋白質核酸酵素. 38. 647-657 (1993)

[文献書誌] 加藤尚子 (藤澤久雄): "T3ファージを用いた巨大DNAのクローニングと構造解析" 蛋白質核酸酵素「ゲノム解析研究」. 38. 618-625 (1993)