1991 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	03221105
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	加納康正京都薬科大学, 生命薬学研究所, 助教授 (50093399) 和田明京都大学, 理学部, 助手 (80025387) 藤田信之国立遺伝学研究所, 分子遺伝, 助手 (90173434) 椙崎弘幸京都大学, 化学研究所, 助手 (60135598) 水野猛名古屋大学, 農学部, 教授 (10174038)
キーワード	大腸菌デ-タベ-ス / 塩基配列決定の高速化 / 湾曲DNA / 転写関始シグナル / 塩基性タンパク質
研究概要	本重点領域研究の計画研究をより円滑に進めるために、一昨年度作製した「大腸菌デ-タベ-ス」の利用プログラムをより一層改良し、Version3.0として班員に配付した。この改良により、各班員がデ-タの訂正を行なうことができるようになり、「大腸菌デ-タベ-ス」がより完全なものになることが期待できる。さらに、新しい塩基配列デ-タ(GenBank70.0)を取り入れ、同時に、各種デ-タの誤りを訂正した。また、塩基配列決定法の改良については、一昨年度来進めてきたトランスポゾンを利用する塩基配列決定法を一段と改良し、PCRを利用しDNAを増幅すること、その際、プライマ-の一端に常磁性のビ-ズを結合させることにより、一本鎖DNAな回収を容易にすること、などの改良を図り、この方法を用いて約80kbの塩基配列決定を行なった。一方、大腸菌ゲノムのもつ各種の構造的ならびに機能的特色についても昨年度にひき続き解析を進めた。その結果、大腸菌のヒストン様タンパク質の遺伝子が自己制御を行なっていること、湾曲DNAに特異的に結合するタンパク質がhns遺伝子の産物であることがわかったこと、定常期のRNAポリメラ-ゼが対数増殖期のものと異なっていること、塩基性タンパク質についてのN末端のアミノ酸配列決定を進め、数種の新しいタンパク質を見出したことなどの成果を挙げている。

研究成果

(23件)

すべてその他

すべて文献書誌 (23件)

[文献書誌] Uemura,Y.: "Cloning,characterization and physical location of the rplY gene which encodes ribosomal protein L25 in Escherichia coli K-12." Mol.Gen.Genet.226. 341-344 (1991)
[文献書誌] Kitakawa,M.: "The mitochondrial ribosome." Biochimie. 73. 813-825 (1991)
[文献書誌] Yura,T.: "Systematic sequencing of the Escherichia coli genome:Analysis of the 0-2.4 min region." Science. (1992)
[文献書誌] Kasai,H.: "Use of a mini-transposon and magnetic beads in PCR-based systematic DNA sequencing of Escherichia coli ordered clones." (1992)
[文献書誌] Jeong,Z.: "Nucleotide sequence and analysis of an Escherichia coli chromosomal region containing the genes for ribosomal proteins L21 and L27." (1992)
[文献書誌] Isono,K.: "Escherichia coli database and its retrieval system:Quick access to genetic and physical maps,ordered clones,nucleotide sequences and related literature." (1992)
[文献書誌] Hiroyuki Sugisaki;Katsuhiko Yamamoto;Mituru Takanami: "The HgaI Restriction-Modification System Contains Two Cytosine Methylase Genes Responsible for Modification of Different DNA Strands" The Journal of Biological Chemistry. 266. 13952-13957 (1991)
[文献書誌] Yamada,H.,et al: "Molecular analysis of the Escherichia coli hns gene encoding a DNA-binding protein,which preferentially recognizus curved DNA sequences." Mol.Gen.Genet.230. 332-337 (1991)
[文献書誌] Tanaka,K.,et al: "Systematic characterization of curved DNA segments randomly cloned from Escherichia coli and their functional significance" Mol.Gen.Genet.226. 367-376 (1991)
[文献書誌] Tanaka,K.et al.: "Overproduction and rapid purification of the Escherichia coli histone-like protein,H-NS." Agric.Biol.Chem.55. 3139-3141 (1991)
[文献書誌] Igarashi,K.,Fujita,N.,& Ishihama,A.: "Identification of a subunit assembly domain in the alpha subunit of Escherichia coli RNA polymerase" Journal of Molecular Biology. 218. 1-8 (1991)
[文献書誌] Nakayama,M.,Fujita,N.,Osawa,S.& Ishihama,A.: "Identification and characterization of RNA polymerase sigma factor from Micrococcus luteus" Journal of Biological Chemistry. 266. 2911-2916 (1991)
[文献書誌] Ozaki,M.,Wada,A.,Fujita,N.,& Ishihama,A.: "Growth phase-dependent modification of RNA polymerase in Escherichia coli" Molecular and General Genetics. 230. 17-23 (1991)
[文献書誌] Ozaki,M.,Fujita,N.,Wada,A.,& Ishihama,A.: "Promoter selectivity of the stationary-phase forms of Escherichia coli RNA polymerase and conversion in vitro of the Sl form enzyme into a log-phase enzyme-like form" Nucleic Acids Research. 20. 257-261 (1992)
[文献書誌] Miyazawa,S.,Makivic,M.,Horiki,T.and Homma,S.: "Numerical Expreiments on Frustrated Quantum Spin Systems" in"Computational Approach in Condensed Matter Phenomena",Tokyo:Springer Verlag. (1992)
[文献書誌] Miwako Ozaki;Akira Wada;Nobuyuki Fujita & Akira Ishihama: "Growth phase-dependent modification of RNA polymerase in Escherichia coli" Mol Gen Gent. 230. 24-27 (1991)
[文献書誌] Miwako Ozaki,Nobuyuki Fujita,Akira Wada & Akira Ishihama: "Promoter selectivity of the stationary-phase forms of Escherichia coli RNA polymerase and conversion in vitro of the Sl form enzyme into a log-phase enzyme-like form" Nucleic Acids Research. 20. 257-261 (1992)
[文献書誌] Kano,Y.: "Participation of the histone-like protein HU and of IHF in minichromosomal maintenance in Escherichia coli." Gene. 103. 25-30 (1991)
[文献書誌] Kenri,T.: "Construction and characterization of an Escherichia coli mutant with a deletion of the metZ gene encoding tRNA_<f1>^<met>." Gene. 103. 31-36 (1991)
[文献書誌] Kenri,T.: "Construction and characterization of an Escherichia coli mutant deficient in the metY gene encoding tRNA^<Met>_<f2>:either tRNA^<Met>_<f1> or ^<Met>_<f2> is required for cell growth" Gene.
[文献書誌] Kohno,K.: "Autogenous regulation of transcription of the hupA gene in Escherichia coli" Proc.Natl.Acad.Sci.USA.
[文献書誌] Goshima,N.: "HU-2 Mutants of Escherichia coli deficient in DNA binding"
[文献書誌] Goshima,N.: "Increased ability of DNA-binding of HU-2 by the amino acid substitution in the carboxy-terminal arm domain."

1991 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] Uemura,Y.: "Cloning,characterization and physical location of the rplY gene which encodes ribosomal protein L25 in Escherichia coli K-12." Mol.Gen.Genet.226. 341-344 (1991)

[文献書誌] Kitakawa,M.: "The mitochondrial ribosome." Biochimie. 73. 813-825 (1991)

[文献書誌] Yura,T.: "Systematic sequencing of the Escherichia coli genome:Analysis of the 0-2.4 min region." Science. (1992)

[文献書誌] Kasai,H.: "Use of a mini-transposon and magnetic beads in PCR-based systematic DNA sequencing of Escherichia coli ordered clones." (1992)

[文献書誌] Jeong,Z.: "Nucleotide sequence and analysis of an Escherichia coli chromosomal region containing the genes for ribosomal proteins L21 and L27." (1992)

[文献書誌] Isono,K.: "Escherichia coli database and its retrieval system:Quick access to genetic and physical maps,ordered clones,nucleotide sequences and related literature." (1992)

[文献書誌] Hiroyuki Sugisaki;Katsuhiko Yamamoto;Mituru Takanami: "The HgaI Restriction-Modification System Contains Two Cytosine Methylase Genes Responsible for Modification of Different DNA Strands" The Journal of Biological Chemistry. 266. 13952-13957 (1991)

[文献書誌] Yamada,H.,et al: "Molecular analysis of the Escherichia coli hns gene encoding a DNA-binding protein,which preferentially recognizus curved DNA sequences." Mol.Gen.Genet.230. 332-337 (1991)

[文献書誌] Tanaka,K.,et al: "Systematic characterization of curved DNA segments randomly cloned from Escherichia coli and their functional significance" Mol.Gen.Genet.226. 367-376 (1991)

[文献書誌] Tanaka,K.et al.: "Overproduction and rapid purification of the Escherichia coli histone-like protein,H-NS." Agric.Biol.Chem.55. 3139-3141 (1991)

[文献書誌] Igarashi,K.,Fujita,N.,& Ishihama,A.: "Identification of a subunit assembly domain in the alpha subunit of Escherichia coli RNA polymerase" Journal of Molecular Biology. 218. 1-8 (1991)

[文献書誌] Nakayama,M.,Fujita,N.,Osawa,S.& Ishihama,A.: "Identification and characterization of RNA polymerase sigma factor from Micrococcus luteus" Journal of Biological Chemistry. 266. 2911-2916 (1991)

[文献書誌] Ozaki,M.,Wada,A.,Fujita,N.,& Ishihama,A.: "Growth phase-dependent modification of RNA polymerase in Escherichia coli" Molecular and General Genetics. 230. 17-23 (1991)

[文献書誌] Ozaki,M.,Fujita,N.,Wada,A.,& Ishihama,A.: "Promoter selectivity of the stationary-phase forms of Escherichia coli RNA polymerase and conversion in vitro of the Sl form enzyme into a log-phase enzyme-like form" Nucleic Acids Research. 20. 257-261 (1992)

[文献書誌] Miyazawa,S.,Makivic,M.,Horiki,T.and Homma,S.: "Numerical Expreiments on Frustrated Quantum Spin Systems" in"Computational Approach in Condensed Matter Phenomena",Tokyo:Springer Verlag. (1992)

[文献書誌] Miwako Ozaki;Akira Wada;Nobuyuki Fujita & Akira Ishihama: "Growth phase-dependent modification of RNA polymerase in Escherichia coli" Mol Gen Gent. 230. 24-27 (1991)

[文献書誌] Miwako Ozaki,Nobuyuki Fujita,Akira Wada & Akira Ishihama: "Promoter selectivity of the stationary-phase forms of Escherichia coli RNA polymerase and conversion in vitro of the Sl form enzyme into a log-phase enzyme-like form" Nucleic Acids Research. 20. 257-261 (1992)

[文献書誌] Kano,Y.: "Participation of the histone-like protein HU and of IHF in minichromosomal maintenance in Escherichia coli." Gene. 103. 25-30 (1991)

[文献書誌] Kenri,T.: "Construction and characterization of an Escherichia coli mutant with a deletion of the metZ gene encoding tRNA_<f1>^<met>." Gene. 103. 31-36 (1991)

[文献書誌] Kenri,T.: "Construction and characterization of an Escherichia coli mutant deficient in the metY gene encoding tRNA^<Met>_<f2>:either tRNA^<Met>_<f1> or ^<Met>_<f2> is required for cell growth" Gene.

[文献書誌] Kohno,K.: "Autogenous regulation of transcription of the hupA gene in Escherichia coli" Proc.Natl.Acad.Sci.USA.

[文献書誌] Goshima,N.: "HU-2 Mutants of Escherichia coli deficient in DNA binding"

[文献書誌] Goshima,N.: "Increased ability of DNA-binding of HU-2 by the amino acid substitution in the carboxy-terminal arm domain."

1991 年度実績報告書

磯野克己神戸大学, 理学部, 教授 (70011640)