1998 年度実績報告書

ゲノム機能解析

研究課題

研究課題/領域番号	08283105
研究機関	国立遺伝学研究所
研究代表者	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)
研究分担者	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)
キーワード	ボディマップ / EST / ATAC-PCR / 完全長cDNA / 遺伝子破壊 / 遺伝子発現パターン / RNAi / 酵母two-hybrid法
研究概要	成果報告ヒトcDNA解析については、基本臓器35及び広汎な病理組織から3万のEST(GS)を追加し、ボディマップの充実を進めた(大久保)。ATAC-PCR(アダプター付加競合PCR法)の改良を進め、最大1000倍のdynamic rangeをもつ高精度の発現パターン情報の集積を開始した(大久保、加藤)。オリゴキャップ法による高品質の完全長cDNAライブラリーを作成し、神経芽細胞腫、回腸粘膜、大腸粘膜から計7000クローンの5'端の情報を得た(菅野)。127種類の新規完全長クローンの全長塩基配列を決定した(加藤)。未知遺伝子の機能解析のため30種のノックアウトマウスを作成した(高橋)。モデル生物ゲノムの解析では、ます線虫cDNAのカタログ化を全遺伝子の約半数(10000)にまで進めた。うち約3500について発現細胞(系譜)を決定し発現パターンマップデータベースの構築を進めるとともに、RNAiによる系統的機能解析をおこなった(小原)。細胞性粘菌では14000のEST解析を終了し、発現パターン解析法の確立と機能解析法の確立を進めた(前田)。出芽酵母では、遺伝子発現ネットワーク解明をめざして遺伝子破壊株および発現制御株約170を作製した。全長ORFをクローン化した酵母two-hybrid用コンストラクトの約8000株バンク作成し、プーリング戦略による体系的なタンパク相互作用データ収集を開始した(伊藤)。第6染色体の詳細な機能解析も続けた(村上)。枯草菌では、日本の領域の新規遺伝子1000の変異株バンク作成をほぼ終了した。変異株651株のうち38%に表現型を見いだした。さらに556 Kbの領域に合計186の転写単位を同定した(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進めている。

研究成果
(18件)

すべてその他

すべて文献書誌 (18件)

[文献書誌] Matsuo,N.: "A novel SCG10-related gene uniquely expressed in the nervous system." Gene. 215. 477-481 (1998)
[文献書誌] Takada,T.: "Expression of Green Fluorescent Protein in Transgenic Mice." Methods in Enzymology. (in press).
[文献書誌] Yoshida,K.: "cDNA cloning and chromosomal mapping of mouse BH-protocadherin." DNA sequence. (in press).
[文献書誌] Yoshida,K: "Cloning,expression analysis,and chromosomal localization of BH-protocadherin,a novel member of the cadherin superfamily." Genomics. 49. 458-461 (1998)
[文献書誌] Sugano,S.: "Transmembrane-domain trapping:a novel method for isolation of cDNAs encoding putative membrane proteins." DNA Res.5. 187-193 (1998)
[文献書誌] N.-S.Kim: "Cloning of human polyubiquitin cDNAs and a ubiquitin-binding assay involving its in vitro translation product." J.Biochem.124. 35-39 (1998)
[文献書誌] F.Osaka,: "A new NEDD8-ligating system for cullin-4A." Genes Dev.12. 2263-2268 (1998)
[文献書誌] The C.elegans Sequencing Consortium:"Genome sequence of the nematode C.elegans:A platform for investigating biology." Science. 282. 2012-2018 (1998)
[文献書誌] Tabara,H.: "Pos-1 encodes a cytoplasmic zinc-finger protein essential for germline specification in C.elegans." Development. 126. ,1-11 (1999)
[文献書誌] Kuwahara,M.: "A water channel of the nematode Caenorhabditis elegans and its implications for channel selectivity of MIP proteins." The Am.J.Physiol.:Cell Physiol.(in press). (1999)
[文献書誌] Morio,T.: "The dictyostelium developmental cDNA project:Generation and analysis of expressed sequence tags from the first-finger stage of development." DNA Research. 5. 335-340 (1998)
[文献書誌] 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)
[文献書誌] Choi,D-K.: "Fluorescent differential display analysis of gene expression in apoptotic neuroblastoma cells." Gene. 223. 21-31 (1998)
[文献書誌] Ito,T.: "Fluorescent differential display:a fast and reliable method for message display polymerase chain reaction." Methods Enzymol.303. 298-309 (1999)
[文献書誌] 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)
[文献書誌] Shiratori,A.: "Systematic Identification,Classification,and Chracterization of the Open Reading Frames which Encode Novel Helicase-related Proteins in Saccharomyces cerevisiae by Gene Disruption and Northern Analydsis." Yeast. (in press). (1998)
[文献書誌] Moriya,S.: "A Bacillus subtilis gene-encoding protein homologous to eukaryotic SMC motor protein is necessary for chromosome partition." Mol Microbiol,. 29. 179-187 (1998)
[文献書誌] Nanamiya,H.: "ClpC regulates the fate of a sporulation initiation sigma factor,sigmaH protein,in Bacillus subtilis at elevated temperatures." Mol Microbiol,. 29. 505-513 (1998)

1998 年度 実績報告書

ゲノム機能解析

研究代表者

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)

研究成果

[文献書誌] Matsuo,N.: "A novel SCG10-related gene uniquely expressed in the nervous system." Gene. 215. 477-481 (1998)

[文献書誌] Takada,T.: "Expression of Green Fluorescent Protein in Transgenic Mice." Methods in Enzymology. (in press).

[文献書誌] Yoshida,K.: "cDNA cloning and chromosomal mapping of mouse BH-protocadherin." DNA sequence. (in press).

[文献書誌] Yoshida,K: "Cloning,expression analysis,and chromosomal localization of BH-protocadherin,a novel member of the cadherin superfamily." Genomics. 49. 458-461 (1998)

[文献書誌] Sugano,S.: "Transmembrane-domain trapping:a novel method for isolation of cDNAs encoding putative membrane proteins." DNA Res.5. 187-193 (1998)

[文献書誌] N.-S.Kim: "Cloning of human polyubiquitin cDNAs and a ubiquitin-binding assay involving its in vitro translation product." J.Biochem.124. 35-39 (1998)

[文献書誌] F.Osaka,: "A new NEDD8-ligating system for cullin-4A." Genes Dev.12. 2263-2268 (1998)

[文献書誌] The C.elegans Sequencing Consortium:"Genome sequence of the nematode C.elegans:A platform for investigating biology." Science. 282. 2012-2018 (1998)

[文献書誌] Tabara,H.: "Pos-1 encodes a cytoplasmic zinc-finger protein essential for germline specification in C.elegans." Development. 126. ,1-11 (1999)

[文献書誌] Kuwahara,M.: "A water channel of the nematode Caenorhabditis elegans and its implications for channel selectivity of MIP proteins." The Am.J.Physiol.:Cell Physiol.(in press). (1999)

[文献書誌] Morio,T.: "The dictyostelium developmental cDNA project:Generation and analysis of expressed sequence tags from the first-finger stage of development." DNA Research. 5. 335-340 (1998)

[文献書誌] 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)

[文献書誌] Choi,D-K.: "Fluorescent differential display analysis of gene expression in apoptotic neuroblastoma cells." Gene. 223. 21-31 (1998)

[文献書誌] Ito,T.: "Fluorescent differential display:a fast and reliable method for message display polymerase chain reaction." Methods Enzymol.303. 298-309 (1999)

[文献書誌] 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)

[文献書誌] Shiratori,A.: "Systematic Identification,Classification,and Chracterization of the Open Reading Frames which Encode Novel Helicase-related Proteins in Saccharomyces cerevisiae by Gene Disruption and Northern Analydsis." Yeast. (in press). (1998)

[文献書誌] Moriya,S.: "A Bacillus subtilis gene-encoding protein homologous to eukaryotic SMC motor protein is necessary for chromosome partition." Mol Microbiol,. 29. 179-187 (1998)

[文献書誌] Nanamiya,H.: "ClpC regulates the fate of a sporulation initiation sigma factor,sigmaH protein,in Bacillus subtilis at elevated temperatures." Mol Microbiol,. 29. 505-513 (1998)

1998 年度実績報告書

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)