大腸菌ゲノムの一次構造の解析

1990 Fiscal Year Annual Research Report

• Project/Area Number: 02237105
• Research Institution: Kobe University
• Principal Investigator: 磯野 克己 神戸大学, 理学部, 教授 (70011640)
• Co-Investigator(Kenkyū-buntansha): 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) ; 山岸 正裕 国立遺伝学研究所, 分子遺伝学, 助手 (00220252) ; 水野 毅 名古屋大学, 農学部, 教授 (10174038) ; 椙崎 弘幸 京都大学, 化学研究所, 助手 (60135598)
• Keywords: 大腸菌デ-タベ-ス / 塩基配列決定の高速化 / 湾曲DNA / 転写開始シグナル / 塩基性タンパク質

Research Abstract

本重点領域研究の計画研究をより円滑に進めるために、昨年度作成した「大腸菌デ-タベ-ス」の利用プログラムを大幅に改良するとともに、新しい塩基配列デ-タ(GenBank65.0)と新しい遺伝子地図デ-タ(Bachmann,1990)を取り入れ、Version2.0として完成させ班員に配付した。また、塩基配列決定法の改良については、昨年度来進めているトランスポゾンを利用しての塩基配列決定の高速化を、ほぼ実用化できる段階まで進めることができた。それとともに、いかに塩基対の個数を正確に読み取るかという方法の確立を目指し、必要なコンピュ-タ-ソフトの改良を行なった。これらの結果、従来の方法の3〜5倍程度の速度で塩基配列決定を進めることができる見通しがついた。
一方、大腸菌ゲノムの各種の構造的ならびに機能的特色についても解析を進めた。昨年度に引き続き大腸菌ゲノム中にある湾曲DNAの同定と構造解析を行なうとともに、湾曲DNAに結合する蛋白質を見出した。これによって湾曲DNAの持つ生理的意義の解析への糸口が見出された。また、大腸菌ゲノム中の転写開始シグナル・プロモ-タ-の分布と強度について解析をすすめるとともに、これまであまり解析の進んでいない塩基性蛋白質についても、改良2次元電気泳動による包括的解析を進めることができた。

Research Products

• [Publications] Oh,B.ーK.: "The location of a gene for a small stable(10Sa)RNA in the Escherichia coli Chromosome." J.Bacteriol.172. 4708-4709 (1990)
• [Publications] Uemura,Y.: "Cloning,characterization and physical location of the rplY gene which encodes ribosomal protein L25 in Escherichia coli Kー12." Mol.Gen.Genet.(1991)
• [Publications] Isono,K.: "The Escherichia coli database and its retrieval system: Quick access to genetic and physical map,ordered clones,nucleotide sequences and related literature." (1991)
• [Publications] Jeong,Z.: "Nucleotide sequence and analysis of a region of Escherichia coli chromosome containing the genes for ribosomal proteins L21 and L27." (1991)
• [Publications] Hiroyuki Sugisaki: "The <Hga>___ーI RestrictionーModification System Contains Two Cytosine Methylase Genes Responsible for Modification of Different DNA Strands" Journal of Biological Chemistry.
• [Publications] Muramatsu,S.: "Nucleotide sequence of the region encompassing the <int>___ー gene of a cryptic prophage and <dnaY>___ー gene flanked by a curved DNA sequence of <K.coli>___ー Kー12" Mole.gen.Genet.220. 325-328 (1990)
• [Publications] Yamada,H.: "An <Escherichia coli>___ー protein that preferentially binds tp sharply curved DNA" J.Biochem.(Tokyo). 108. 420-425 (1990)
• [Publications] Tanaka,K.: "Systematic characterization of curved DNA segments randomaly colned from <Escherichia coli>___ー and their functional significance." Mol.gen.Genet.(1991)
• [Publications] Igarashi,K.: "Sequence analysis of two temperatureーsensitive mutations in the alpha subunit gene(rpoA)of Escherichia coli RNA polymerase." Nucleic Acids Res.18. 5945-5948 (1990)
• [Publications] Kobayashi,M.: "Promoter selectivity of Escherichia coli RNA polymerase:Effect of base substitutions in the promoterー35 region on promoter strength." Nucleic Acids Res.18. 7367-7372 (1990)
• [Publications] Igarashi,K.: "Identification of a subunit assembly domain in the alpha subunit of Escherichia coli RNA polymerase." J.Mol.Biol.(1991)
• [Publications] Ishihama,A.: "Molecular mechanisms of transcription regulation: Promoter selectivity of RNA polymerase." Proc.5th FAOB Cong.(1991)
• [Publications] Miyauchi,T.: "Basigen,a new,broadly distributed member of the immunoglobulin superfamily,has strong homology with both the immunoglobulin V domain and betaーchain of major histocompatibility complex class II antigen." J.Biochem.107. 316-323 (1990)
• [Publications] Miyazawa,S.: "Estimation of the average energy increment by amino acid exchange in proteins and its use in evaluating a homology scoring matrix." Protein Engineering. 3. 318-318 (1990)
• [Publications] Miyazawa,S.: "A new substitution matrix for protein sequence searches based on contact frequencies in crystal."
• [Publications] Miyazawa,S.: "Estimation of protein stability changes for single site substitution mutants."
• [Publications] 宮澤 三造: "ゲノム解析とデ-タベ-ス" 実験医学. 8. 42-48 (1990)
• [Publications] 宮澤 三造: "核酸デ-タベ-スの現状と将来" 現代化学. 239. 38-43 (1991)
• [Publications] AKIRA WADA: "Structure and probable genetic location of a“ribosome modulation factor"associated with 100S ribosomes in stationaryーphase Escherichia coli cells" Proc.Natl.Acad.Sci.USA. 87. 2657-2661 (1990)
• [Publications] Yasunobu Kano: "Requirement of integration host factor (IHF) for growth of Escherichia coli deficient in HU protein." Gene. 89. 133-137 (1990)
• [Publications] Naoki Goshima: "HUー1 mutants of Escherichia coli Deficient in DNA binding." Gene. 96. 141-145 (1990)
• [Publications] Naoki Goshima: "Characterization of HUーlile protein from Bifidobacterium longum." Biochimie. 72. 207-212 (1990)
• [Publications] Yohei Natori: "Genetic transfornmation of Lactobacillus casei by electroporation." Biochimie. 72. 265-269 (1990)
• [Publications] Kyoko Kohno: "Promoters and autogeneous control of the escherichia coli hupA and hupB genes." J.Mol.Biol.213. 27-36 (1990)
• [Publications] Yohei Natori: "Nucleotide sequences and genomic constitution of five tryptophan genes of Lactobacillus casei." J.Biochem. 107. 248-255 (1990)
• [Publications] Teru Ogura: "Maintenance of plasmids in HU and IHF mutants of Escherichia coli." Molec.Gen.Genet.220. 197-203 (1990)
• [Publications] Fumio Imamoto: "Physiological characterization of deletion mutants of the hupA and hupB genes in Escherichia coli." The Bacterial Chromosome ed.by Karl Drlica and Monica Riley American Society for Microbiology,Washington,DC. 259-266 (1990)
• [Publications] 石浜 明: "“バイオサイエンス戦略マニュアル"(瀬野悍二ら編)" 共立出版, 13 (1990)
• [Publications] Ishihama,A.: "“Structure and Function of Nucleic Acids and Proteins"(CーW.Wu and F.Y.ーH.Wu,eds)" Raven Press,New York, 8 (1990)
• [Publications] Ishihama,A.: "“Control of Cell Growth and Division"(A.Ishihama and H.Yoshikawa,eds.)" Jpn.Sci.Press/SpringerーVerlag,