1996 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
研究種目	重点領域研究
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, 構造遺伝学研究センター, 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 / 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
研究概要	成果報告ヒトcDNA解析については、まずヒト各組織40およびマウス主要臓器20から3′無作為シーケンスを約60000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701種類の新規蛋白質遺伝子を含む1284種類の完全長cDNAクローンを同定し、82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、マウスを用いたジーンターゲティング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約半数(7000)にまで進め、約1000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)。出芽酵母ゲノムでは第6染色体の遺伝子破壊実験の結果127のORFのうち増殖必須遺伝子は24個であり新規のものは7個であった。これらについて5つの条件でノーザン解析をおこなった(村上)。また12の破壊株に蛍光ディファレンシャルディスプレイ法を応用し発現変化のみられるバンドを多数同定した。さらにゲノム塩基配列からバンド同定ができるよう全遺伝子を網羅するプライマーのセットをデザインするとともに、系統的2ハイブリッド法を開発した(伊藤)。枯草菌ゲノムではわが国で決定された1.4Mb領域の約1400の遺伝子のうち240の遺伝子の破壊変異株を作成したところ、23個で表現型が見出され、2個の2成分制御系遺伝子が見つかった。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のシステムの準備をした(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
(26件)

すべてその他

すべて文献書誌 (26件)

[文献書誌] Tabara H.: "A multi-well version of in situ hybridization on whole mount embryos of C.elegans." Nucl.Acids Res.24. 2119-2124 (1996)
[文献書誌] Nishida K.: "An expression profile of active genes in human corneal epithelium and the isolation of human keratin 12 cDNA." Invest Ophthalmol Vis Sci.37. 1800-1809 (1996)
[文献書誌] Ohno I.: "A cDNA cloning of Human AEBP1 from Primary Cultured Osteoblast and Its Expression in a Differentiating Osteoblastic Cell Line." B.B.R.C.228. 411-414 (1996)
[文献書誌] Shimizu-Matsumoto A.: "Isolation and chromosomal localization of human cone cGMP phosphodiesteraseg cDNA." Genomics.32. 121-124 (1996)
[文献書誌] Kita H.: "An expression profile of active genes in cultured human keratinocytes." DNA research.3. 1-7 (1996)
[文献書誌] Maeda K.: "cDNA cloning and expression of a novel adipose specific collagen-like factor,apM1 (AdiPose Most abundant Gene transcript 1)." Biochem Biophys Res Commun.221. 286-289 (1996)
[文献書誌] Okubo K.: "Complementary DNA sequence (EST) collections and the expression information of the human genome." FEBS letters.(in press). (1997)
[文献書誌] Tsurumi C.: "cDNA cloning and functional analysis of the p97 subunit of the 26S proteasome,a polypeptide identical to the type-1 tumor-necrosis-factor-receptor-associated protein-2/55.11." Eur.J.Biochem.239. 912-921 (1996)
[文献書誌] Yamaguchi T.: "Cloning and expression of cDNA encoding a human ubiquitin-conjugating enzyme similar to the Drosophila bendless gene product." J.Biochem.120. 494-497 (1996)
[文献書誌] Ozaki K.: "Cloning,expression,and chromosomal mapping of a novel cyclophilin-related gene (PPIL 1) from human fetal brain." Cytogenet Cell Genet. 72. 242-245 (1996)
[文献書誌] Watanabe T.K.: "Cloning,expression,and mapping of TCTEL 1,a putative human homologue of murine Tcte 1,to 6q." Cytogenet Cell Genet. 73. 153-156 (1996)
[文献書誌] Watanabe T.K.: "Cloning,expression pattern and mapping to 12p13.2->p13.1 of CLAPS3,a gene encoding a novel clathrin-adaptor small chain." Cytogenet Cell Genet. 73. 214-217 (1996)
[文献書誌] Ito T.: "An SH3 domain-mediated interaction between the phagocyte NADPH oxidase factors p40phox and p47phox" FEBS Lett.385. 229-232 (1996)
[文献書誌] Sumimoto H.: "Assembly and activation of the phagocyte NADPH oxidase : specific interaction of the N-terminal Src Homology 3 (SH3) domain of p47phox with p22phox is required for activation of the NADPH oxidase" J.Biol.Chem.271. 22152-22158 (1996)
[文献書誌] Kito K.: "Fluorescent differential display analysis of gene expression in differentiating neuroblastoma cells" Gene. 184. 73-81 (1997)
[文献書誌] Furihata C.: "Rare occurrence of ras and p53 gene mutation in mouse stomach tumors induced by N-methyl-N-nitrosourea" Jpn.J.Cancer Res.in press.(1997)
[文献書誌] Furihata C.: "Differentially expressed MHC class II-associated invariant chain in rat stomach pyloric mucosa with N-methyl-N'-nitro-N-nitrosoguanidine exposure" Cancer Res.in press. (1997)
[文献書誌] Naitou M.: "Sequencing of 23kb fragment,clone gapB and 4682 of Saccharomyces cerevisiae chromosome VI." Yeast. 12. 77-84 (1996)
[文献書誌] Eki T.: "Analysis of a 36.2-kb DNA sequence including the right telomere of chromosome VI from Saccharomyces cerevisiae" Yeast. 12. 149-167 (1996)
[文献書誌] Eki T.: "Fifteen open readig frames in a 30.8-kb region of the right arm of chromosome VI from Saccharomyces cerevisiae" Yeast. 12. 177-190 (1996)
[文献書誌] A.Goffeau: "Life with 6000 genes" Science. 274. 546-567 (1996)
[文献書誌] Murakami Y.: "Systematic analysis of novel genes of the Saccharomyces cerevisiae chromosome VI" Microbial and Comparative Genomics. 3. 244 (1996)
[文献書誌] Ogasawara N.: "The systematic sequencing of the Bacillus Subtilis genome in Japan" Microbiol.142. 2993-2994 (1996)
[文献書誌] Yamamoto H.: "Determination of a 12kb nucleotide sequence around the 76 degree region of the Bacillus subtilis chromosome" Microbiol.142. 1417-1421 (1996)
[文献書誌] Sadaie Y.: "The nucleotide sequence of the phoB-rrnE-groESL region of the Bacillus subtilis chromosome" Microbiol.(in press). (1997)
[文献書誌] Kasahara Y.: "Sequence analysis of the 36Kb region between gntZ and trnY genes of Bacillus subtilis genome" DNA Res.(in press). (1997)

1996 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, 構造遺伝学研究センター, 教授 (70135292)

研究成果

[文献書誌] Tabara H.: "A multi-well version of in situ hybridization on whole mount embryos of C.elegans." Nucl.Acids Res.24. 2119-2124 (1996)

[文献書誌] Nishida K.: "An expression profile of active genes in human corneal epithelium and the isolation of human keratin 12 cDNA." Invest Ophthalmol Vis Sci.37. 1800-1809 (1996)

[文献書誌] Ohno I.: "A cDNA cloning of Human AEBP1 from Primary Cultured Osteoblast and Its Expression in a Differentiating Osteoblastic Cell Line." B.B.R.C.228. 411-414 (1996)

[文献書誌] Shimizu-Matsumoto A.: "Isolation and chromosomal localization of human cone cGMP phosphodiesteraseg cDNA." Genomics.32. 121-124 (1996)

[文献書誌] Kita H.: "An expression profile of active genes in cultured human keratinocytes." DNA research.3. 1-7 (1996)

[文献書誌] Maeda K.: "cDNA cloning and expression of a novel adipose specific collagen-like factor,apM1 (AdiPose Most abundant Gene transcript 1)." Biochem Biophys Res Commun.221. 286-289 (1996)

[文献書誌] Okubo K.: "Complementary DNA sequence (EST) collections and the expression information of the human genome." FEBS letters.(in press). (1997)

[文献書誌] Tsurumi C.: "cDNA cloning and functional analysis of the p97 subunit of the 26S proteasome,a polypeptide identical to the type-1 tumor-necrosis-factor-receptor-associated protein-2/55.11." Eur.J.Biochem.239. 912-921 (1996)

[文献書誌] Yamaguchi T.: "Cloning and expression of cDNA encoding a human ubiquitin-conjugating enzyme similar to the Drosophila bendless gene product." J.Biochem.120. 494-497 (1996)

[文献書誌] Ozaki K.: "Cloning,expression,and chromosomal mapping of a novel cyclophilin-related gene (PPIL 1) from human fetal brain." Cytogenet Cell Genet. 72. 242-245 (1996)

[文献書誌] Watanabe T.K.: "Cloning,expression,and mapping of TCTEL 1,a putative human homologue of murine Tcte 1,to 6q." Cytogenet Cell Genet. 73. 153-156 (1996)

[文献書誌] Watanabe T.K.: "Cloning,expression pattern and mapping to 12p13.2->p13.1 of CLAPS3,a gene encoding a novel clathrin-adaptor small chain." Cytogenet Cell Genet. 73. 214-217 (1996)

[文献書誌] Ito T.: "An SH3 domain-mediated interaction between the phagocyte NADPH oxidase factors p40phox and p47phox" FEBS Lett.385. 229-232 (1996)

[文献書誌] Sumimoto H.: "Assembly and activation of the phagocyte NADPH oxidase : specific interaction of the N-terminal Src Homology 3 (SH3) domain of p47phox with p22phox is required for activation of the NADPH oxidase" J.Biol.Chem.271. 22152-22158 (1996)

[文献書誌] Kito K.: "Fluorescent differential display analysis of gene expression in differentiating neuroblastoma cells" Gene. 184. 73-81 (1997)

[文献書誌] Furihata C.: "Rare occurrence of ras and p53 gene mutation in mouse stomach tumors induced by N-methyl-N-nitrosourea" Jpn.J.Cancer Res.in press.(1997)

[文献書誌] Furihata C.: "Differentially expressed MHC class II-associated invariant chain in rat stomach pyloric mucosa with N-methyl-N'-nitro-N-nitrosoguanidine exposure" Cancer Res.in press. (1997)

[文献書誌] Naitou M.: "Sequencing of 23kb fragment,clone gapB and 4682 of Saccharomyces cerevisiae chromosome VI." Yeast. 12. 77-84 (1996)

[文献書誌] Eki T.: "Analysis of a 36.2-kb DNA sequence including the right telomere of chromosome VI from Saccharomyces cerevisiae" Yeast. 12. 149-167 (1996)

[文献書誌] Eki T.: "Fifteen open readig frames in a 30.8-kb region of the right arm of chromosome VI from Saccharomyces cerevisiae" Yeast. 12. 177-190 (1996)

[文献書誌] A.Goffeau: "Life with 6000 genes" Science. 274. 546-567 (1996)

[文献書誌] Murakami Y.: "Systematic analysis of novel genes of the Saccharomyces cerevisiae chromosome VI" Microbial and Comparative Genomics. 3. 244 (1996)

[文献書誌] Ogasawara N.: "The systematic sequencing of the Bacillus Subtilis genome in Japan" Microbiol.142. 2993-2994 (1996)

[文献書誌] Yamamoto H.: "Determination of a 12kb nucleotide sequence around the 76 degree region of the Bacillus subtilis chromosome" Microbiol.142. 1417-1421 (1996)

[文献書誌] Sadaie Y.: "The nucleotide sequence of the phoB-rrnE-groESL region of the Bacillus subtilis chromosome" Microbiol.(in press). (1997)

[文献書誌] Kasahara Y.: "Sequence analysis of the 36Kb region between gntZ and trnY genes of Bacillus subtilis genome" DNA Res.(in press). (1997)

1996 年度実績報告書

小原雄治国立遺伝学研究所, 構造遺伝学研究センター, 教授 (70135292)