ゲノム機能解析

1997 Fiscal Year Annual Research Report

• Project/Area Number: 08283105
• Research Institution: National Institute of Genetics
• Principal Investigator: 小原 雄治 国立遺伝学研究所, 生物遺伝資源情報総合センター, 教授 (70135292)
• Co-Investigator(Kenkyū-buntansha): 加藤 誠志 (財)相模中央化学研究所, 主任研究員 ; 小笠原 直毅 奈良先端科学技術大学院大学, 教授 (10110553) ; 村上 康文 理化学研究所, 筑波研究センター, 先任研究員 (90200279) ; 伊藤 隆司 東京大学, 医科学研究所, 助手 (90201326) ; 大久保 公策 大阪大学, 細胞生体工学センター, 助教授 (40233069)
• Keywords: ボディマップ / cDNA / 遺伝子破壊 / 遺伝子発現パターン / 線虫 / 出芽酵母 / 枯草菌 / ディファレンシャルディスプレイ

Research Abstract

成果報告
ヒトcDNA解析については、まずヒト各組織54およびマウス主要臓器から3'無作為シーケンスを約110,000集積しボディマップの充実を進めた(大久保)。また完全長cDNAの解析を進め1260種類の新規蛋白質遺伝子を含む2130種類の完全長cDNAクローンを同定し、200種類の新規完全長の配列決定をした(加藤)。未知遺伝子の機能解析のために酵母、マウスを用いた遺伝子破壊システムの確立をおこなった(高橋)。モデル生物ゲノムの解析では、線虫ゲノムのcDNAのカタログ化を全遺伝子の半数以上の8000種にまで進め、約2000種について発現細胞(系譜)を決定し発現パターンマップの構築を進めた(小原)。細胞性粘菌のcDNA解析を進めるため、18,000クローンをグリッド化し、タグ塩基配列決定、アンチセンスRNA発現コンストラクト作成を進めた(前田)。出芽酵母ゲノムでは第6染色体の遺伝子破壊実験から見いだされた24個の増殖必遺伝子について表現型解析をおこなった(村上)。EUグループとの共同で野生株、破壊株、過剰産生株の間でFDD解析を行い、新規の標的遺伝子候補22種及び8種の遺伝子を負の標的遺伝子として同定した。酵母全ORFの間でtwo-hybrid解析による蛋白質相互作用解析の準備をした(伊藤)。枯草菌ゲノムではわが国で決定された14Mb領域の約1400の遺伝子のうち443の遺伝子の破壊変異株を作成した。表現型のシステマチックな検出のために、7種類のC源の利用能、カタボライト感受性、蛋白質の分泌能、細胞の運動性、DNA損傷の感受性、塩・エタノール・過酸化水素・温度などのストレスに対する感受性、SDSに対する感受性を調べるための21種類のプレートを選び、コロニーの大きさ(増殖能)とLacZ活性による呈色の変化(発現)を調べる作業を開始した。また全蛋白の2次元電気泳動での同定と破壊株との対応付けを進めた(小笠原)。以上の研究結果は東大医科研ヒトゲノムセンターなどにおいてインターネット上での公開をおこなっている。

Research Products

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• [Publications] Sadaie,Y.: "Nucleotide sequence and analysis of the phoB-rrnE-groESL region of the Bacillus subtilis chromosome." Microbiology. 143. 1861-1866 (1997)
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• [Publications] Hata,K.: "Anionic amphiphile-independent activation of the phagocyte NADPH oxidase in a cell-free system by p47phox and p67phox,both in C terminally truncated forms:lmplication for regulatory SH3 domain-mediated interactions" J.Biol.Chem.273. 4232-4236 (1998)
• [Publications] Kato,K.: "Adaptor-tagged competitive PCR: a novel m ethod for measuring relative gene expression." Nucleic Acids Research. 25. 4694-4696 (1997)
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• [Publications] Shimizu,F.: "Isolation of a novel human cDNA (rho HP1) homologous to rho genes." Biochemica et Biophysica Acta. 1351. 13-16 (1997)
• [Publications] Takahashi,N.: "Molecular propaties of the proteasome activator PA28 family proteins and gamma-interferon regulation." Genes to Cells. 2. 195-211 (1997)
• [Publications] Watanabe,K.T.: "Cloning and characterization of a novel member of the human Mad gene family (MADH)." Genomics. 42. 446-451 (1997)
• [Publications] Maeda K: "Analysis of an expression profile of genes in the human adipose tissue" Gene. 190. 227-235 (1997)
• [Publications] Okubo K: "Complementary DNA sequence(EST)collections and the expression information of the human genome" FFBS letters. 403. 225-229 (1997)
• [Publications] Matsumoto A: "An expression profile of Genes in Human Retina and Isolation of a cDNA for a Novel Rod Photoreceptor protein" IOVS. 38. 2576-2585 (1997)
• [Publications] Kuriyama H: "Molecular cloning and Expression of a Novel Aquaporin from Adipose Tissue with Glycerol permeabllity" BBRC. 241. 53-58 (1997)
• [Publications] Matsuo N: "Cloning of a cDNA encoding a novel sugar transporter expressed in the neonatal mouse hippocampus" BBRC. 238. 126-129 (1997)
• [Publications] Blazejewski S: "Expression in human liver and cultured human liver myofibroblasts" Am.j.of Pathology. 151. 651-657 (1997)
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• [Publications] Takei,Y.: "Molecular cloning of a novel gene similar to myeroid antigen CD33 and its specific expression in placenta." Cytogenet.Cell Genet.78. 295-300 (1997)
• [Publications] Omori,Y.: "Cloning and mapping of a novel human zinc-finger gene TCF-17 homologous to rodent kid1." Cytogenet.Cell Genet.78. 285-288 (1997)
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• [Publications] Hill,A.: "Assignment of fragile site 8E(FRA8E) of human chromosome band 8q24.11 adjacent to the hereditary multiple exostoses 1 gene and two overlapping Langer-Giedeon syndrome deletion endpoints." Cytogenet. Cell Genet.78. 56-57 (1997)
• [Publications] Eki,T.: "Cloning and characterization of novel gene, DCRR1, expressed from Down's syndrome critical region of human chromosome 21q22.2" DNA Sequence. 7. 153-164 (1997)
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• [Publications] Naitou,M.: "Expression profiles of transcripts from 126 ORFs in Saccharomyces cerevisiae chromosome VI by systematic Northern analyse" Yeast. (in press). (1997)
• [Publications] Ishiai,M.: "Isolation of human and fission yeast gene homologues of the budding yeast origin recognition complex subunit Orc5:Human homologue (PRC5L) maps to chromosome band 7q22" Genomics. (in press).
• [Publications] Eki,T.: "Mapping of the human genes encoding cyclin H (CCNH) and CDK-activating kinase (CAK) assembly factor MAT1 (MNAT1) to chromosome bands 5q13.3-q14 and 14q23, respectively," Genomics. (in press).
• [Publications] Goffeau.A.: "Yeast Genome Directory" Nature. Supplement to Vol 387,29 May. (1997)
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• [Publications] Nakazato,A: "Comparison of the structural characteristics of chromosome VI in Saccharomyces sensu stricto:The divergence, species-dependent features and uniqueness of Sake yeast" Yeast. (in press). (1998)
• [Publications] 村上康文: "ヒト遺伝子全容の理解を目指して:その1 ゲノム解析研究の目的とその方法" 科学技術ジャーナル. 1月号. 66-69 (1998)
• [Publications] 村上康文: "ヒト遺伝子全容の理解を目指して:その2 ゲノム解析研究の目的とその方法" 科学技術ジャーナル. 2月号. 66-69 (1998)
• [Publications] 村上康文: "ヒト遺伝子全容の理解を目指して:その3 塩基配列の解読法" 科学技術ジャーナル. 3月号. 66-69 (1998)
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• [Publications] Mizuki,K.: "Functional modules and expression of mouse p40phox and p67phox, Src homology 3 (SH3) domain-containing proteins involved in the phagocyte NADPH oxidase complex" Eur.J.Biochem.251. 573-582 (1998)
• [Publications] Nakamura,R.: "The PC motif:A novel and evolutionarily conserved sequence involved in interaction between p40phox and p67phox,SH3 domain-containing cytosolic factors of the phagocyte NADPH oxidase" Eur.J.Biochem.251. 583-589 (1998)
• [Publications] 村上康文: "酵母の機能解析プロジェクト" 蛋白質核酸酵素. 42巻(17). 3067-3069 (1997)
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• [Publications] L.Aubry: "The Dictyostelium mitogen-activated protein kinase ERK2 is regulated by ras and cAMP-dependent protein kinase (PKA) and mediates PKA function." J.Bio.Chem.vol.272(7). 3883-3886 (1997)
• [Publications] 小原雄治: "線虫(1000細胞のシンフォニ-)" 共立出版株式会社, 1-153 (1997)