1992 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	04NP0401
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	大木操埼玉県立がんセンター研究所, 部長 (00158792) 中村祐輔癌研究会, 癌研究所, 部長 (70217909) 吉田光昭東京大学, 医科学研究所, 教授 (80012607) 吉川寛大阪大学, 医学部, 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た。これらの成果は、ゲノムデータベース作成において1991年度に引き続き、国際的成果の約半分近くを占める貢献である。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。これまで2年間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。 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13000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。本年度を以て良質な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の全長解析などを可能にするいろいろな技術開発が行われた。 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、多色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、BUDRを用いる染色体の分染とin situハイブリッド法の組み合わせ技術の確立など、本班の成果を問う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とマッピング作業の成果が出始める時代が近づいたと思われる。

研究成果

(49件)

すべてその他

すべて文献書誌 (49件)

[文献書誌] Ichikawa,H(Ohki,M): "Long-Distance restriction mapping of the proximal long arm of human chromosome 21 with NotI linking clones" Proc.Natl.Acad.Sci.USA. 89. 23-27 (1992)
[文献書誌] Emi,M(Nakamura,Y): "Isolation and mapping of 88 new RFLP markers on human chromosome 8" Genomics. 13. 1261-1266 (1992)
[文献書誌] Hattori,M(Sakaki,Y): "Orphan peak analysis:A novel method for detection of point mutations using an automated fluorescent DNA sequencer" Genomics.
[文献書誌] Kawasaki,K(Shimizu,N): "Shotgun Polymerase Chain Reaction:Construction of Clone Libraries Specific to a NotI Fragment of Flow-Sorted Human Chromosome 22" Genomics. 13(1). 109-114 (1992)
[文献書誌] Tabe,H(Tsuji,S): "Isolation of X chromosome:Specific cDNA clones expressed in human brain" Am J Hum Genet. 51. A402 (1992)
[文献書誌] Nagafuchi,S(Nakagome,Y): "A majority of the marker chromosome in Japanese patients with Turner syndrome are derived from Y chromosomes" Hum.Genet. 89. 590-592 (1992)
[文献書誌] Iizuka,M(Hayashi,K): "Cloning and polymerase chain reaction:Single strand conformation polymorphism analysis of anonymous Alu repeats on chromosome 11" Genomics. 12. 139-146 (1992)
[文献書誌] Matsumoto,K(Inoko,H): "Cluster of fibronectin type III repeat found in the human major histocompatibility complex class III region shows the highest homology with the repeats in an extracellular matrix protein,tenascin" Genomics. 12. 485-491 (1992)
[文献書誌] Matsuda,F: "Structure and physical map of 64 Variable segments in the 3'0.8-megabase region of the human immunoglobulin heavy-chain locus" Nature Genetics. 3. 88-94 (1993)
[文献書誌] Chifu,Y(Imamura,T): "β-Thalassemia major resulting from a compound heterozygosity for the β-globin gene mutation:further evidence for multiple origin and migration of the thalassemia gene" Hum Genet. 89. 343-346 (1992)
[文献書誌] Muro,Y(Masumoto,H): "Centromere protein B assembles human centromeric alpha-satellite DNA at the 17 bp sequence,CENP-B box" J.Cell Biol. 116. 585-596 (1992)
[文献書誌] 藤山秋佐夫: "デュアルレーザー染色体ソーティングとその利用" 蛋白質核酸酵素増刊「ゲノム解析研究」. 38. 586-590 (1993)
[文献書誌] Isobe,M: "Localization of the gene encoding the α subunit of human interleukin-5 receptor(IL5RA)to chromosome region 3p24-3p26" Genomics. 14. 755-758 (1992)
[文献書誌] Hirota,T(Niikawa,N): "Microdissection of human chromosomal regions 8q23.3-q24.11 and 2q33-qter:Construction of DNA libraries and isolation of their clones" Genomics. 13. 349-354 (1992)
[文献書誌] NIH/CEPH collaborative mapping group(Nakamura,Y): "A comprehensive genetic linkage map of the human genome" Science. 258. 67-86 (1992)
[文献書誌] Schellenberg,G.D.(Miki,T): "Homozygosity mapping and Werner's syndrome" Lancet. 339. 1002 (1992)
[文献書誌] Sano,A(Kondo,I): "Human dopamin transpoter gene polymorphism in Japanese population" Hum Genet.
[文献書誌] Wakisaka,A: "Linkage analysis of hereditary spinocerebellar ataxias." Molecular approaches to the study and treatment of human diseases. 9-15 (1992)
[文献書誌] Suzuki,T(Yoshida,M): "A trans-activator Tax of human T cell leukemia virus type 1(HTLV-1)interacts with cAMP-responsive element(CRE)binding protein and CRE modulator proteins that bind to the 21-base-pair enhancer of HTLV-1" Proc.Natl.Acad.Sci.USA. 89. 610-614 (1993)
[文献書誌] Sugano,S: "Use of epitope-tagged cDNA library to isolate cDNAs encoding proteins with nuclear localization potential" Gene. 120. 227-233 (1992)
[文献書誌] Okubo,K: "Large scale cDNA sequencing for analysis of quantative and qualitative aspects of gene expression" Nature genetics. 2. 173-179 (1992)
[文献書誌] Nomura,N: "Prediction of new human genes by entire sequencing of randomly sampled cDNA clones"
[文献書誌] Hara,E(Oda,K): "Id-related genes encoding HLH proteins are required for G1 progression and are repressed in senescent human fibroblasts"
[文献書誌] Hirashima,H(Shimada,K): "Nucleotide sequence of the third cytokine LD78 gene and mapping of all three LD78 gene loci to human chromosome 17" DNA Sequence. (1993)
[文献書誌] Takayanagi,A(Ayusawa,D): "Intron 1 an the 5'-flanking region of the human thymidylate snthase gene as a regulatory determinnt of growth-dependent expression" Nucleic Acid Res. 20. 4021-4025 (1992)
[文献書誌] Miyajima,A(Arai,K): "Cytokine receptors and signal transduction" Ann.Rev.Immunol. 10. 295-331 (1992)
[文献書誌] Nakayama,T: "Structural change of a particular H2B histone gene possibly results in differences in its transcriptional regulation in different chicken tissues" Nucleic Acids Res. 20. 1309-1314 (1992)
[文献書誌] Shibuya,H(Taniguchi,T): "IL-2 and EGF Receptors Stimulate the Hematopoietic Cell Cycle via Different Signaling Pathways:Demonstration of a Novel Role for c-myc" Cell. 70. 57-67 (1992)
[文献書誌] Seto,Y(Nagata,S): "Chromosomal gene organization colony-stimulating factor receptor" J.Immunol. 148. 259-266 (1992)
[文献書誌] Eguchi,Y: "Prediction of the Functional Sites in Amino Acid Sequences" Genome Informatics Workshop III. 77-80 (1992)
[文献書誌] Minagawa,K(Doi,M): "Change of the higer order structure of DNA induced by the complexation with intercalating synthetic polymer,as is visualized by fluorescence microscopy" Nucleic Acids Research. 21. 37-40 (1993)
[文献書誌] Shikata,T(Kotaka,T): "Biased sinusoidal field gel electrophoresis for size-dependent DNA separation" Biopolymers. 31. 253-254 (1991)
[文献書誌] Kido,K(Otsuka,E): "Sequence-dependent cleavage of DNA by alkylation with antisense oligodeoxyribonucleotides containing a 2-(N-iodo-acetylaminoethyl)thio-adenine" Nucleic Acids Res. 20. 1339-1344 (1992)
[文献書誌] Ichikawa,A(Sugiura,Y): "Activation of DNA Cleavage by Dynemicin A in a B-Z Conformational Junction" Biochemistry. 31. 6784-6787 (1992)
[文献書誌] Hashimoto,C(Fujisawa,H): "DNA sequences necessary for packaging of bacteriophage T3 DNA" Virology. 187. 788-796 (1992)
[文献書誌] 河野享子(和田守正): "YACクローンの安定化" 蛋白質核酸酵素. 38. 607-612 (1993)
[文献書誌] Tokino,T(Imai,T): "Physical mapping of a 950-kb region surrounding a locus(D10S102)tightly linked to the MEN2A gene" Genomics. 12. 394-400 (1992)
[文献書誌] Ito,W(Kurosawa,Y): "Development of a prokaryote expression system that exploits dicistronic gene organization" Gene. 118. 87-91 (1992)
[文献書誌] Hayashizaki,Y(Mukai,T): "A new method for constructing Not I linking and cutting library using restrictiontrapper" Genomics. 14. 733-739 (1992)
[文献書誌] Takahashi,E: "A high-resolution cytogenetic map of human chromosome 3:localization of 291 new cosmid markers by direct R-banding fluorescence in situ hybridization" Genomics. 13. 1047-1055 (1992)
[文献書誌] Yoshida,M: "Recent advances in fluorescence in situ hybridization" J.Radiat.Res. 33. 54-60 (1992)
[文献書誌] Naito,M(Kohara,Y): "Identification of a homeobox-containing gene located between lin-45 and unc-24 on chromosome IV in the nematode Caenorhabditis elegans" Nucleic Acid Res. 20. 2967-2969 (1992)
[文献書誌] Ogasawara,N: "Genes and their organization in replication origin region of bacterial chromosome" Mol.Microbiol. 6. 629-634 (1992)
[文献書誌] Yokota,H(Oishi,M): "Differential cloning of genomic DNA:Cloning of DNA with an altered primary stucture by in-gel competitive reassociation" Proc.Natl.Aca.Sci.USA. 87. 6398-6402 (1990)
[文献書誌] Baba,Y: "Preparation of Polyacrylamide Gel Filled Capillaries for Ultrahigh Resolution of Polynucleotides by Capillary Gel Electrophoresis" Anal.Chem. 64. 1221-1225 (1992)
[文献書誌] S.Selig(Okumura,K): "Delineation of DNA Replication Time Zones by Fluorscecce in situ Hybridization" EMBO J. 11. 1217-1225 (1992)
[文献書誌] 陶山明: "OHFデータから整列クローン・ライブラリーを求める方法" 情報処理学基礎. 27. 57-62 (1992)
[文献書誌] Inazawa,J(Abe,T): "Assignment of human Fas gene to chromosome 10 at q24.1" Genomics. 14. 821-822 (1992)
[文献書誌] N.Fujiki: "Session of international opinion suvery" Huamn Genome Research and Society. 119-165 (1992)

1992 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] Ichikawa,H(Ohki,M): "Long-Distance restriction mapping of the proximal long arm of human chromosome 21 with NotI linking clones" Proc.Natl.Acad.Sci.USA. 89. 23-27 (1992)

[文献書誌] Emi,M(Nakamura,Y): "Isolation and mapping of 88 new RFLP markers on human chromosome 8" Genomics. 13. 1261-1266 (1992)

[文献書誌] Hattori,M(Sakaki,Y): "Orphan peak analysis:A novel method for detection of point mutations using an automated fluorescent DNA sequencer" Genomics.

[文献書誌] Kawasaki,K(Shimizu,N): "Shotgun Polymerase Chain Reaction:Construction of Clone Libraries Specific to a NotI Fragment of Flow-Sorted Human Chromosome 22" Genomics. 13(1). 109-114 (1992)

[文献書誌] Tabe,H(Tsuji,S): "Isolation of X chromosome:Specific cDNA clones expressed in human brain" Am J Hum Genet. 51. A402 (1992)

[文献書誌] Nagafuchi,S(Nakagome,Y): "A majority of the marker chromosome in Japanese patients with Turner syndrome are derived from Y chromosomes" Hum.Genet. 89. 590-592 (1992)

[文献書誌] Iizuka,M(Hayashi,K): "Cloning and polymerase chain reaction:Single strand conformation polymorphism analysis of anonymous Alu repeats on chromosome 11" Genomics. 12. 139-146 (1992)

[文献書誌] Matsumoto,K(Inoko,H): "Cluster of fibronectin type III repeat found in the human major histocompatibility complex class III region shows the highest homology with the repeats in an extracellular matrix protein,tenascin" Genomics. 12. 485-491 (1992)

[文献書誌] Matsuda,F: "Structure and physical map of 64 Variable segments in the 3'0.8-megabase region of the human immunoglobulin heavy-chain locus" Nature Genetics. 3. 88-94 (1993)

[文献書誌] Chifu,Y(Imamura,T): "β-Thalassemia major resulting from a compound heterozygosity for the β-globin gene mutation:further evidence for multiple origin and migration of the thalassemia gene" Hum Genet. 89. 343-346 (1992)

[文献書誌] Muro,Y(Masumoto,H): "Centromere protein B assembles human centromeric alpha-satellite DNA at the 17 bp sequence,CENP-B box" J.Cell Biol. 116. 585-596 (1992)

[文献書誌] 藤山 秋佐夫: "デュアルレーザー染色体ソーティングとその利用" 蛋白質核酸酵素増刊「ゲノム解析研究」. 38. 586-590 (1993)

[文献書誌] Isobe,M: "Localization of the gene encoding the α subunit of human interleukin-5 receptor(IL5RA)to chromosome region 3p24-3p26" Genomics. 14. 755-758 (1992)

[文献書誌] Hirota,T(Niikawa,N): "Microdissection of human chromosomal regions 8q23.3-q24.11 and 2q33-qter:Construction of DNA libraries and isolation of their clones" Genomics. 13. 349-354 (1992)

[文献書誌] NIH/CEPH collaborative mapping group(Nakamura,Y): "A comprehensive genetic linkage map of the human genome" Science. 258. 67-86 (1992)

[文献書誌] Schellenberg,G.D.(Miki,T): "Homozygosity mapping and Werner's syndrome" Lancet. 339. 1002 (1992)

[文献書誌] Sano,A(Kondo,I): "Human dopamin transpoter gene polymorphism in Japanese population" Hum Genet.

[文献書誌] Wakisaka,A: "Linkage analysis of hereditary spinocerebellar ataxias." Molecular approaches to the study and treatment of human diseases. 9-15 (1992)

[文献書誌] Suzuki,T(Yoshida,M): "A trans-activator Tax of human T cell leukemia virus type 1(HTLV-1)interacts with cAMP-responsive element(CRE)binding protein and CRE modulator proteins that bind to the 21-base-pair enhancer of HTLV-1" Proc.Natl.Acad.Sci.USA. 89. 610-614 (1993)

[文献書誌] Sugano,S: "Use of epitope-tagged cDNA library to isolate cDNAs encoding proteins with nuclear localization potential" Gene. 120. 227-233 (1992)

[文献書誌] Okubo,K: "Large scale cDNA sequencing for analysis of quantative and qualitative aspects of gene expression" Nature genetics. 2. 173-179 (1992)

[文献書誌] Nomura,N: "Prediction of new human genes by entire sequencing of randomly sampled cDNA clones"

[文献書誌] Hara,E(Oda,K): "Id-related genes encoding HLH proteins are required for G1 progression and are repressed in senescent human fibroblasts"

[文献書誌] Hirashima,H(Shimada,K): "Nucleotide sequence of the third cytokine LD78 gene and mapping of all three LD78 gene loci to human chromosome 17" DNA Sequence. (1993)

[文献書誌] Takayanagi,A(Ayusawa,D): "Intron 1 an the 5'-flanking region of the human thymidylate snthase gene as a regulatory determinnt of growth-dependent expression" Nucleic Acid Res. 20. 4021-4025 (1992)

[文献書誌] Miyajima,A(Arai,K): "Cytokine receptors and signal transduction" Ann.Rev.Immunol. 10. 295-331 (1992)

[文献書誌] Nakayama,T: "Structural change of a particular H2B histone gene possibly results in differences in its transcriptional regulation in different chicken tissues" Nucleic Acids Res. 20. 1309-1314 (1992)

[文献書誌] Shibuya,H(Taniguchi,T): "IL-2 and EGF Receptors Stimulate the Hematopoietic Cell Cycle via Different Signaling Pathways:Demonstration of a Novel Role for c-myc" Cell. 70. 57-67 (1992)

[文献書誌] Seto,Y(Nagata,S): "Chromosomal gene organization colony-stimulating factor receptor" J.Immunol. 148. 259-266 (1992)

[文献書誌] Eguchi,Y: "Prediction of the Functional Sites in Amino Acid Sequences" Genome Informatics Workshop III. 77-80 (1992)

[文献書誌] Minagawa,K(Doi,M): "Change of the higer order structure of DNA induced by the complexation with intercalating synthetic polymer,as is visualized by fluorescence microscopy" Nucleic Acids Research. 21. 37-40 (1993)

[文献書誌] Shikata,T(Kotaka,T): "Biased sinusoidal field gel electrophoresis for size-dependent DNA separation" Biopolymers. 31. 253-254 (1991)

[文献書誌] Kido,K(Otsuka,E): "Sequence-dependent cleavage of DNA by alkylation with antisense oligodeoxyribonucleotides containing a 2-(N-iodo-acetylaminoethyl)thio-adenine" Nucleic Acids Res. 20. 1339-1344 (1992)

[文献書誌] Ichikawa,A(Sugiura,Y): "Activation of DNA Cleavage by Dynemicin A in a B-Z Conformational Junction" Biochemistry. 31. 6784-6787 (1992)

[文献書誌] Hashimoto,C(Fujisawa,H): "DNA sequences necessary for packaging of bacteriophage T3 DNA" Virology. 187. 788-796 (1992)

[文献書誌] 河野 享子(和田 守正): "YACクローンの安定化" 蛋白質核酸酵素. 38. 607-612 (1993)

[文献書誌] Tokino,T(Imai,T): "Physical mapping of a 950-kb region surrounding a locus(D10S102)tightly linked to the MEN2A gene" Genomics. 12. 394-400 (1992)

[文献書誌] Ito,W(Kurosawa,Y): "Development of a prokaryote expression system that exploits dicistronic gene organization" Gene. 118. 87-91 (1992)

[文献書誌] Hayashizaki,Y(Mukai,T): "A new method for constructing Not I linking and cutting library using restrictiontrapper" Genomics. 14. 733-739 (1992)

[文献書誌] Takahashi,E: "A high-resolution cytogenetic map of human chromosome 3:localization of 291 new cosmid markers by direct R-banding fluorescence in situ hybridization" Genomics. 13. 1047-1055 (1992)

[文献書誌] Yoshida,M: "Recent advances in fluorescence in situ hybridization" J.Radiat.Res. 33. 54-60 (1992)

[文献書誌] Naito,M(Kohara,Y): "Identification of a homeobox-containing gene located between lin-45 and unc-24 on chromosome IV in the nematode Caenorhabditis elegans" Nucleic Acid Res. 20. 2967-2969 (1992)

[文献書誌] Ogasawara,N: "Genes and their organization in replication origin region of bacterial chromosome" Mol.Microbiol. 6. 629-634 (1992)

[文献書誌] Yokota,H(Oishi,M): "Differential cloning of genomic DNA:Cloning of DNA with an altered primary stucture by in-gel competitive reassociation" Proc.Natl.Aca.Sci.USA. 87. 6398-6402 (1990)

[文献書誌] Baba,Y: "Preparation of Polyacrylamide Gel Filled Capillaries for Ultrahigh Resolution of Polynucleotides by Capillary Gel Electrophoresis" Anal.Chem. 64. 1221-1225 (1992)

[文献書誌] S.Selig(Okumura,K): "Delineation of DNA Replication Time Zones by Fluorscecce in situ Hybridization" EMBO J. 11. 1217-1225 (1992)

[文献書誌] 陶山 明: "OHFデータから整列クローン・ライブラリーを求める方法" 情報処理学基礎. 27. 57-62 (1992)

[文献書誌] Inazawa,J(Abe,T): "Assignment of human Fas gene to chromosome 10 at q24.1" Genomics. 14. 821-822 (1992)

[文献書誌] N.Fujiki: "Session of international opinion suvery" Huamn Genome Research and Society. 119-165 (1992)

1992 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] 藤山秋佐夫: "デュアルレーザー染色体ソーティングとその利用" 蛋白質核酸酵素増刊「ゲノム解析研究」. 38. 586-590 (1993)

[文献書誌] 河野享子(和田守正): "YACクローンの安定化" 蛋白質核酸酵素. 38. 607-612 (1993)

[文献書誌] 陶山明: "OHFデータから整列クローン・ライブラリーを求める方法" 情報処理学基礎. 27. 57-62 (1992)