1989 年度実績報告書

大腸菌ゲノムの一次構造の解析

研究課題

研究課題/領域番号	01655005
研究機関	神戸大学
研究代表者	磯野克己神戸大学, 理学部, 教授 (70011640)
研究分担者	宮澤三造国立遺伝学研究所, 遺伝情報センター, 助教授 (60190774) 藤田信之国立遺伝学研究所, 分子遺伝, 助手 (90173434) 水野猛名古屋大学, 農学部, 教授 (10174038) 橘秀樹神戸大学, 理学部, 助教授 (70126118) 椙崎弘幸京都大学, 化学研究所, 助手 (60135598)
キーワード	整列クローンバンク / 塩基配列決定 / DNAシーケンサー / コンピュータープログラム / 大腸菌データベース
研究概要	大腸菌ゲノム構造の全体像を解明するためには、(1)実験データを効率よく得るための方法・技術を開発すること、及び、(2)データの収録・整理・統合などを効率よく行なうこと、が必要である。(1)の目的のためには、大腸菌の全ゲノムを網羅したいわゆる「整列クローンバンク」から、個々のクローンを取り出して塩基配列を決定する過程、および塩基配列決定を可能な限り機械化・自動化することを検討する必要がある。そこで本研究では、個々のクローンのDNAを直接、塩基配列決定のための酵素反応(ジデオキシ法)の鋳型として用いる方法を開発し、これを市販されているDNAシーケンサーで使用できるようにした。さらに、DNAシーケンサーをよりよく駆使できるように、ハード、ソフトの両面から改良を加えた。また、(2)の目的のためにはコンピューターの使用が不可欠であり、これを最大限有効に利用する方法を講じなければならない。大腸菌ゲノムの全構造が解明されれば、その約470万塩基対からなる塩基配列データは、それ自身で一個の独立のデータベースとして、取り扱われるようになるものと考えられる。そこで、すでに発表されている塩基配列データーを収録して整理し、これに遺伝子地図データおよび整列クローンのデータを加えて「大腸菌データーベース」として整理し、班員の利用に供した。このように本年度の研究では、実験方法の改良、特に実験の機械化、コンピューター化を計ることを綿密に検討するとともに、データの評価法を確立し、研究を進めていく過程でこれらをより普遍的なものに編成し、本重点領域研究の他の計画研究・公募研究を含めた実際の研究に反映させてきた。それとともに、各種の領域の塩基配列決定を行ない、大腸菌のゲノム全構造の解明に向けての努力を重ねた。

研究成果

(21件)

すべてその他

すべて文献書誌 (21件)

[文献書誌] Ueshima R.,Fujita N.,Ishihama A.: "DNA supercoiling and temperature shift affect the promoter activity of the Escherichia coli rpoH gene encoding the heat-shock sigma subunit of RNA rolymerase." Mol.Gen.Genet.215. 185-189 (1989)
[文献書誌] Nakayama M.,Fujita N.,Ohama T.,Osawa S.,Ishihama A.: "Micrococcus luteus,a bacterium with a high G+C content,contains Escherichia coli-type promoters." Mol.Gen.Genet.218. 384-389 (1989)
[文献書誌] Tanaka Y.,Tsujimura A.,Fujita N.,Isono S.,Isono K.: "Cloning and analysis of an Escherichia coli operon containing the rpmF gene for ribosomal protein L32 and the gene for a 30-kilodalton protein." J.Bacteriol.171. 5707-5712 (1989)
[文献書誌] Igarashi K.,Fujita N.,Ishihama A.: "Promoter selectivity of Escherichia coli RNA polymerase:omega factor is responsible for the ppGpp sensitivity." Nucleic Acids Res.17. 8755-8765 (1989)
[文献書誌] Wada A.,Yamazaki Y.,Fujita N.,Ishihama A: "Identification and probable genetic location of the ribosome modulation factor associated with 100S ribosomes in stationaryphase Escherichia coli cells." Proc.Natl.Acad.Sci.U.S.A.87. (1990)
[文献書誌] Igarashi K.,Fujita N.,Ishihama A.: "Functional mapping of alpha subunit of Escherichia coli RNA polymerase."
[文献書誌] Kang,W.-K.,Icho T.,Isono S.,Kitakawa M.,Isono K.: "Characterization of the gene rimK responsible for the addition of glutamic acid residues to the C-terminus of ribosomal protein S6 in Escherichia coli K12." Mol.Gen.Genet.217. 281-288 (1989)
[文献書誌] Tanaka S.,Matsushita Y.,Yoshikawa A.,Isono K.: "Cloning and molecular characterization of the gene rimL which encodes an enzyme acetylating ribosomal protein L12 of Escherichia coli." Mol.Gen.Genet.217. 289-293 (1989)
[文献書誌] Matsushita Y.,Kitakawa M.,Isono K.: "Cloning and analysis of the nuclear genes for two mitochondrial ribosomal proteins in yeast." Mol.Gen.Genet.219. 119-124 (1989)
[文献書誌] Tanaka Y.,Tujimura A.,Fujita N.,Isono S.,Isono K.: "Cloning and analysis of an Escherichia coli operon containing the rpmF gene for ribosomal protein L32 and the gene for a 30 kilodalton protein." J.Bacteriol.171. 5707-5712 (1989)
[文献書誌] Kitakawa M.,Isono K.,Grohmann L.,Graack.H.-R.: "Cloning and characterization of nuclear genes for two mitochondrial ribosomal proteins in Saccharomyces cerevisiae." Nucl.Acids Reds.(1990)
[文献書誌] Yoshikawa A.,Isono K.: "Chromosome III of Saccharomyces cerevisiae:An ordered clone bank,a detailed restriction map and analysis of transcripts suggests the presence of 160 genes." Yeast. (1990)
[文献書誌] Oh,B.-K.,Isono K.,Apirion,D.: "THE location of a gene for a small stable(10Sa)RNA in the Escherichia coli choromosome." J.Bacteriol.(1990)
[文献書誌] Miyazawa S.: "DNA data bank of Japan: Present status and future plans." Computers and DNA, SFI Studies in the Sciences of Complexity. VII. 47-61 (1990)
[文献書誌] Miyauchi T.: "Basigi,a new member of the immunoglobulin superfamily of broad distribution has strong homology with both immunoglobulin V domain and the β-chain of major histocompatibility complex class II antigen." J.Biochem.(1990)
[文献書誌] Muramatsu S.,Mizuno T.: "Nucleotide sequence of the region encompassing the glpFKF operon and its upstream region containing a bent DNA sequence of E.coli." Nucleic Acids Research. 17. 1898 (1989)
[文献書誌] Muramatsu S.,Mizuno T.: "Nucleotide Sequence of the fabE gene and flanking regions containing a bent DNA sequence of E.coli." Nucleic Acids Research. 17. 3982 (1989)
[文献書誌] Muramatsu S.,Mizuno T.: "Nucleotide sequence of the region encompassing the int gene of a cryptic prophage and the dnaY gene flanked by a curved DNA sequence of E.coli Kー12" Mol Gen Genet. 220. 325-328 (1990)
[文献書誌] K.Kita,H.Kotani,H.Sugisaki & M.Takanami: "The FokI Restriction-Modification System I.Organization and Nucleotide Sequences of the Restriction and Modification Genes." The Journal of Biological Chemistry. 264. 5751-5756 (1989)
[文献書誌] H.Sugisaki,K.Kita & M.Takanami: "The FokI Restriction-Modification System II.Presence of Two Domains in FokI Methylase Responsible for Modification of Different DNA Strands." The Journal of Biological Chemistry. 264. 5757-5761 (1989)
[文献書誌] Hideki Tachibana: "An 'initiator-terminator vector'suitable for direct expression of genic segments in Escherichia coli" Protein Engineering. (1990)

1989 年度 実績報告書

大腸菌ゲノムの一次構造の解析

研究代表者

磯野 克己 神戸大学, 理学部, 教授 (70011640)

研究成果

[文献書誌] Ueshima R.,Fujita N.,Ishihama A.: "DNA supercoiling and temperature shift affect the promoter activity of the Escherichia coli rpoH gene encoding the heat-shock sigma subunit of RNA rolymerase." Mol.Gen.Genet.215. 185-189 (1989)

[文献書誌] Nakayama M.,Fujita N.,Ohama T.,Osawa S.,Ishihama A.: "Micrococcus luteus,a bacterium with a high G+C content,contains Escherichia coli-type promoters." Mol.Gen.Genet.218. 384-389 (1989)

[文献書誌] Tanaka Y.,Tsujimura A.,Fujita N.,Isono S.,Isono K.: "Cloning and analysis of an Escherichia coli operon containing the rpmF gene for ribosomal protein L32 and the gene for a 30-kilodalton protein." J.Bacteriol.171. 5707-5712 (1989)

[文献書誌] Igarashi K.,Fujita N.,Ishihama A.: "Promoter selectivity of Escherichia coli RNA polymerase:omega factor is responsible for the ppGpp sensitivity." Nucleic Acids Res.17. 8755-8765 (1989)

[文献書誌] Wada A.,Yamazaki Y.,Fujita N.,Ishihama A: "Identification and probable genetic location of the ribosome modulation factor associated with 100S ribosomes in stationaryphase Escherichia coli cells." Proc.Natl.Acad.Sci.U.S.A.87. (1990)

[文献書誌] Igarashi K.,Fujita N.,Ishihama A.: "Functional mapping of alpha subunit of Escherichia coli RNA polymerase."

[文献書誌] Kang,W.-K.,Icho T.,Isono S.,Kitakawa M.,Isono K.: "Characterization of the gene rimK responsible for the addition of glutamic acid residues to the C-terminus of ribosomal protein S6 in Escherichia coli K12." Mol.Gen.Genet.217. 281-288 (1989)

[文献書誌] Tanaka S.,Matsushita Y.,Yoshikawa A.,Isono K.: "Cloning and molecular characterization of the gene rimL which encodes an enzyme acetylating ribosomal protein L12 of Escherichia coli." Mol.Gen.Genet.217. 289-293 (1989)

[文献書誌] Matsushita Y.,Kitakawa M.,Isono K.: "Cloning and analysis of the nuclear genes for two mitochondrial ribosomal proteins in yeast." Mol.Gen.Genet.219. 119-124 (1989)

[文献書誌] Tanaka Y.,Tujimura A.,Fujita N.,Isono S.,Isono K.: "Cloning and analysis of an Escherichia coli operon containing the rpmF gene for ribosomal protein L32 and the gene for a 30 kilodalton protein." J.Bacteriol.171. 5707-5712 (1989)

[文献書誌] Kitakawa M.,Isono K.,Grohmann L.,Graack.H.-R.: "Cloning and characterization of nuclear genes for two mitochondrial ribosomal proteins in Saccharomyces cerevisiae." Nucl.Acids Reds.(1990)

[文献書誌] Yoshikawa A.,Isono K.: "Chromosome III of Saccharomyces cerevisiae:An ordered clone bank,a detailed restriction map and analysis of transcripts suggests the presence of 160 genes." Yeast. (1990)

[文献書誌] Oh,B.-K.,Isono K.,Apirion,D.: "THE location of a gene for a small stable(10Sa)RNA in the Escherichia coli choromosome." J.Bacteriol.(1990)

[文献書誌] Miyazawa S.: "DNA data bank of Japan: Present status and future plans." Computers and DNA, SFI Studies in the Sciences of Complexity. VII. 47-61 (1990)

[文献書誌] Miyauchi T.: "Basigi,a new member of the immunoglobulin superfamily of broad distribution has strong homology with both immunoglobulin V domain and the β-chain of major histocompatibility complex class II antigen." J.Biochem.(1990)

[文献書誌] Muramatsu S.,Mizuno T.: "Nucleotide sequence of the region encompassing the glpFKF operon and its upstream region containing a bent DNA sequence of E.coli." Nucleic Acids Research. 17. 1898 (1989)

[文献書誌] Muramatsu S.,Mizuno T.: "Nucleotide Sequence of the fabE gene and flanking regions containing a bent DNA sequence of E.coli." Nucleic Acids Research. 17. 3982 (1989)

[文献書誌] Muramatsu S.,Mizuno T.: "Nucleotide sequence of the region encompassing the int gene of a cryptic prophage and the dnaY gene flanked by a curved DNA sequence of E.coli Kー12" Mol Gen Genet. 220. 325-328 (1990)

[文献書誌] K.Kita,H.Kotani,H.Sugisaki & M.Takanami: "The FokI Restriction-Modification System I.Organization and Nucleotide Sequences of the Restriction and Modification Genes." The Journal of Biological Chemistry. 264. 5751-5756 (1989)

[文献書誌] H.Sugisaki,K.Kita & M.Takanami: "The FokI Restriction-Modification System II.Presence of Two Domains in FokI Methylase Responsible for Modification of Different DNA Strands." The Journal of Biological Chemistry. 264. 5757-5761 (1989)

[文献書誌] Hideki Tachibana: "An 'initiator-terminator vector'suitable for direct expression of genic segments in Escherichia coli" Protein Engineering. (1990)

1989 年度実績報告書

磯野克己神戸大学, 理学部, 教授 (70011640)