1992 年度実績報告書

形質転換と大量集積

研究課題

研究課題/領域番号	04252102
研究機関	名古屋大学
研究代表者	町田泰則名古屋大学, 理学部, 教授 (80175596)
研究分担者	佐藤文彦京都大学, 農学部, 助教授 (10127087) 中村研三名古屋大学, 農学部, 教授 (80164292) 内宮博文東京大学, 応用微生物研究所, 教授 (50142229) 岩渕雅樹京都大学, 理学部, 教授 (30000839)
キーワード	Tiプラスミドベクター / ヒストン遺伝子 / スポラシン遺伝子 / ショ糖誘導遺伝子 / rolC遺伝子 / シス領域 / ベルベリン合成 / ACPハイドロラーゼ
研究概要	本年度は、以下のような成果を挙げた。 (1)Agrobacteniumによる植物細胞の形質転換に関与している二つのタンパク質を同定し、それらの直接的な相互作用により、形質転換反応が誘発されることがわかった。また、タバコの根特異的プロモーターを用いることにより、スコポラミン生産能力が増大した形質転換ベラドンナの毛状根を作出した。その蓄積量はCaMV35Sプロモーターを用いた場合より、1.5倍高かった。(2)ヒストンH3遺伝子の細胞周期でのS期特異的発現は、二つのcis配列の協調的作用によることがわかった。(3)ショ糖に応答するrolCプロモーター領域に結合する一本鎖結合タンパク質を同定した。(4)ショ糖に応答するスポラミン遺伝子プロモーターには正と負のcis配列が存在することがわかった。(5)ベルベリン生合成の2つの0メチル化酵素のcDNAをクローン化した。(6)イオウ代謝サイクルに中心的な役割を果たしているシステイン合成酵素で細胞質と葉緑体に存在するものの遺伝子cDNAを高等植物からクローニングした。(7)エリシターによるイソフラボノイド生合成の発現とカルコン合成酵素遺伝子の大腸菌での発現を行った。(8)グルタミン合成酵素遺伝子の発現部位が、維管束細胞であることがわかった。(9)植物ウィルスRNA複製酵素遺伝子を発現している植物細胞を利用した物質生産系を確立した。(10)脂肪酸代謝の鍵酵素であるアシルACPヒドロラーゼの33kDaポリペプチドのアミノ末端の配列を34残基決定した。以上のようにこの3年間で、物質生産を担っている植物遺伝子のクローニングとその発現の分子機構の研究が進むと同時に、実際に遺伝子操作により高いレベルの物質生産系が確立しうるところまで進んできた。以上の研究を遂行するために、計画通り設備備品を購入した。

研究成果

(40件)

すべてその他

すべて文献書誌 (40件)

[文献書誌] Yasunori Machida: "Molecular interactions between Agrobacterium and plant cells in Molecualr Mechanisms of Bacterial Virulence (ed.by E.W.Nester)Kluwer" 85-96 (1993)
[文献書誌] Takefumi Kawata: "A cDNA Clone encoding HBP-1b homologue in Arabidopsis thaliana" Nucl.Acids Res.20. 1141 (1992)
[文献書誌] Takuya Nakayama: "Highly conserved hexamer,octamer and nonamer motifs are positive cis-regulatory elements of the wheat histone H3 gene" FEBS Lett.300. 167-170 (1992)
[文献書誌] Sumiko Sugaya: "Deletion analysis of the 5'-upstream region of the Agrobacterium rhizogenes Ri plasmid rolC gene required for tissue-specific expression" Plant Physiol.99. 464-467 (1992)
[文献書誌] Akihiro Suzuki: "Signal-strand.DNA of 5'-upstream region of rolC gene interacts with unclear proteins of carrot cell culture" Biochem.&Biophys.Res.Comm.188. 727-733 (1992)
[文献書誌] Sumiko Katayama: "Analysis of intergenic spacer regions in the nuclear rDNA of Pharbitis nil" Genome. 35. 92-97 (1992)
[文献書誌] Taku Takahashi: "Isolation and analysis of the expression of two genes for the 81-KD heat-shock proteins from Arabidopsis" Plant Physiol.99. 383-390 (1992)
[文献書誌] Sumie Ishiguro: "The nuclear factor SP8BF binds to the 5'-upstream regions of three different genes coding for major tuberous root protiens of sweet potato" Plant Mol.Biol.18. 97-108 (1992)
[文献書誌] Masa-aki Ohto: "Induction of expression of genes coding for sporamin and β-amylase by polygalacturonic acid in leaf-petiole cutting of sweet potato" Plant Physiol.99. 422-427 (1992)
[文献書誌] Tomokazu Konishi: "Protein transport into thylakoid lumen-A stromal processing and energy requirement for the import of the precursor for the 23-kDa protein of PS II." Plant Cell Physiol.(1993)
[文献書誌] 渡辺昭: "緑葉の老化とグルタミン合成酵素" Nippon Nogeikagaku Kaishi. 66. 932-935 (1992)
[文献書誌] Satomi Takeda: "Nucleotide sequence of a cDNA for Osmotin-like protein from cultured tobacco cells" Pleant Physiol.97. 844-846 (1991)
[文献書誌] Fumihiko Sato: "S-adenosyl-L-methionine:scoulerine-9-0-methyltransferase from cultured Coptis japonica cells" Phytochem.(1992)
[文献書誌] Hyung-Hwang Park: "Isoflavone Glucosides Exist as Their 6¨-0-Malonyl Esters in Pueraria lobata and Its Cell Suspension Cultures" Chem.Pharm.Bull.40. 1978-1980 (1992)
[文献書誌] Takashi Hakamatsuka: "Isoflavone Dimers from Yeast-Extract-Treated Cell Suspension Cultures of Pueraria lobata" Z.Naturforscheung. 47c. 177-182 (1992)
[文献書誌] Hiroyuki Takano: "Rapid clonal-propagation of Matricaria chamomilla by tissue-cultured shoot primordia" Japanese Jorunal of Breeding. 41. 421-426 (1991)
[文献書誌] Toshifumi Hirata: "Lipid constituents oil bodies in shoot primordia of Matricaria chamomilla" Plant Cell Report. (1993)
[文献書誌] Saito Kazuki: "Molecular cloning and bacterial expression of cDNA encoding a plant cysteine synthase" Proc.Natl.Acad.Sci.USA. 89. 8078-8082 (1992)
[文献書誌] Kazuki Saito: "Transgenic herbicide-resistant Atropa belladonna using an Ri binary vector and inheritance of the transgenic trait" Plant Cell Rep.11. 219-224 (1992)
[文献書誌] Yuji Minami: "Isolation and characterization of two momordins,ribosome-inactivating protiens form the seeds of bitter gourd (Momordica charantia)" Biosci.Biotech.Biochem.56. 1470-1471 (1992)
[文献書誌] Masachika Hrie: "Base specificity of two plant seed ribonucleases from Momordica charantia and Luffa cylindrica" Biosci.Biotech.Biochem.57. (1993)
[文献書誌] Hiroyuki Imai: "Acyl-(acyl-carrier protein)hydrolase from squash cotyledons specific to long-chain fatty acids:purification and characterization" Plant Mol.Biol.20. 199-206 (1992)
[文献書誌] Ikuo Nishida: "The gene and the RNA for the precursor to the plastidlocated glycerol-3-phosphate acyltransferase of Arabidopsis thaliana" Plant Mol.Biol.21. 262-272 (1993)
[文献書誌] Kaoru Suzuki: "Effects of 2,6-dichlorobenzonitrile on differentiation to tracheary elements of isolated mesophylle cells of Zinnia elegans and formation of secondary cell walls" Physiol.Plant. 86. 43-48 (1992)
[文献書誌] Masashi Mori: "Infectivity of plasmids containing brome mosaic virus cDNA linked to the cauliflower mosaic virus 35S RNA promoter" j.Gen.Virology. 72. 243-246 (1991)
[文献書誌] Masahi Mori: "Expression of brome mosaic virus-encoded replicase genes in transgenic tobacco plants" J.Gen.Virology. 73. 169-172 (1992)
[文献書誌] Tsutomu Furuya: "Biotransformation of Tabersonine in Cell Suspension Cultures of Catharunthus roseus" Phytochemistry. 31. 3065-3069 (1992)
[文献書誌] Takafumi Yoshikawa: "Continuous Production of Glycosides by Bioreactor Using Ginseng Hairy Root Cultures" Microbiology and Biothechnology. 38. (1993)
[文献書誌] Koji Mikami: "Regulation of cell cycle-dependent gene expression.In"Control of Plant Gene Expression"(D.P.S.Verma,ed.)" CRC Press,Inc., 18 (1993)
[文献書誌] Takuya Nakayama: "Regulation of wheat histone gene expression.In"Critical Reviews in Plant Science"(B.V.Conger,ed.)" CRC Press,Inc., (1993)
[文献書誌] Hirofumi Uchimiya: "Production of transgenic rice plants by direct gene transfer into protoplasts.In Plant Transformation protocols(ed.L.Fitzmaurice and F.Lin)" Academic Press, (1992)
[文献書誌] Yutaka Oono: "Molecular and cellular analysis of the rol genes of Agrobacterium rhizogenes Ri Plasmid." J.of Plant Research, (1993)
[文献書誌] 米田好文: "「植物の分子生物学」シロイヌナズナの核ゲノム構成" 共立出版, 372 (1992)
[文献書誌] 米田好文: "「開花・結実の分子機構」花芽形成開始機構の分子遺伝学的研究" 秀潤社, 290 (1992)
[文献書誌] 中村研三: "「植物の分子生物学」糖による植物遺伝子の発現制御" 共立出版, 8 (1992)
[文献書誌] 中村研三: "植物におけるショ糖による遺伝子発現制御" 生化学, 10 (1992)
[文献書誌] 谷口研至: "苗条原基誘導法「最新バイオテクノロジー全書2 野菜の組織・細胞培養と増殖」" 秀潤社, 8 (1990)
[文献書誌] 谷口研至: "生長点培養現代化学増刊「植物バイオテクノジーII」" 東京化学同人, 11 (1991)
[文献書誌] Ikuo Nishida: "Current Topics in Plant Physiology Series" American Society of Plant Physiology, (1993)
[文献書誌] 古谷力: "Maruzen Advanced Technology 植物工学" 丸善株式会社, 260 (1993)

1992 年度 実績報告書

形質転換と大量集積

研究代表者

町田 泰則 名古屋大学, 理学部, 教授 (80175596)

研究成果

[文献書誌] Yasunori Machida: "Molecular interactions between Agrobacterium and plant cells in Molecualr Mechanisms of Bacterial Virulence (ed.by E.W.Nester)Kluwer" 85-96 (1993)

[文献書誌] Takefumi Kawata: "A cDNA Clone encoding HBP-1b homologue in Arabidopsis thaliana" Nucl.Acids Res.20. 1141 (1992)

[文献書誌] Takuya Nakayama: "Highly conserved hexamer,octamer and nonamer motifs are positive cis-regulatory elements of the wheat histone H3 gene" FEBS Lett.300. 167-170 (1992)

[文献書誌] Sumiko Sugaya: "Deletion analysis of the 5'-upstream region of the Agrobacterium rhizogenes Ri plasmid rolC gene required for tissue-specific expression" Plant Physiol.99. 464-467 (1992)

[文献書誌] Akihiro Suzuki: "Signal-strand.DNA of 5'-upstream region of rolC gene interacts with unclear proteins of carrot cell culture" Biochem.&Biophys.Res.Comm.188. 727-733 (1992)

[文献書誌] Sumiko Katayama: "Analysis of intergenic spacer regions in the nuclear rDNA of Pharbitis nil" Genome. 35. 92-97 (1992)

[文献書誌] Taku Takahashi: "Isolation and analysis of the expression of two genes for the 81-KD heat-shock proteins from Arabidopsis" Plant Physiol.99. 383-390 (1992)

[文献書誌] Sumie Ishiguro: "The nuclear factor SP8BF binds to the 5'-upstream regions of three different genes coding for major tuberous root protiens of sweet potato" Plant Mol.Biol.18. 97-108 (1992)

[文献書誌] Masa-aki Ohto: "Induction of expression of genes coding for sporamin and β-amylase by polygalacturonic acid in leaf-petiole cutting of sweet potato" Plant Physiol.99. 422-427 (1992)

[文献書誌] Tomokazu Konishi: "Protein transport into thylakoid lumen-A stromal processing and energy requirement for the import of the precursor for the 23-kDa protein of PS II." Plant Cell Physiol.(1993)

[文献書誌] 渡辺 昭: "緑葉の老化とグルタミン合成酵素" Nippon Nogeikagaku Kaishi. 66. 932-935 (1992)

[文献書誌] Satomi Takeda: "Nucleotide sequence of a cDNA for Osmotin-like protein from cultured tobacco cells" Pleant Physiol.97. 844-846 (1991)

[文献書誌] Fumihiko Sato: "S-adenosyl-L-methionine:scoulerine-9-0-methyltransferase from cultured Coptis japonica cells" Phytochem.(1992)

[文献書誌] Hyung-Hwang Park: "Isoflavone Glucosides Exist as Their 6¨-0-Malonyl Esters in Pueraria lobata and Its Cell Suspension Cultures" Chem.Pharm.Bull.40. 1978-1980 (1992)

[文献書誌] Takashi Hakamatsuka: "Isoflavone Dimers from Yeast-Extract-Treated Cell Suspension Cultures of Pueraria lobata" Z.Naturforscheung. 47c. 177-182 (1992)

[文献書誌] Hiroyuki Takano: "Rapid clonal-propagation of Matricaria chamomilla by tissue-cultured shoot primordia" Japanese Jorunal of Breeding. 41. 421-426 (1991)

[文献書誌] Toshifumi Hirata: "Lipid constituents oil bodies in shoot primordia of Matricaria chamomilla" Plant Cell Report. (1993)

[文献書誌] Saito Kazuki: "Molecular cloning and bacterial expression of cDNA encoding a plant cysteine synthase" Proc.Natl.Acad.Sci.USA. 89. 8078-8082 (1992)

[文献書誌] Kazuki Saito: "Transgenic herbicide-resistant Atropa belladonna using an Ri binary vector and inheritance of the transgenic trait" Plant Cell Rep.11. 219-224 (1992)

[文献書誌] Yuji Minami: "Isolation and characterization of two momordins,ribosome-inactivating protiens form the seeds of bitter gourd (Momordica charantia)" Biosci.Biotech.Biochem.56. 1470-1471 (1992)

[文献書誌] Masachika Hrie: "Base specificity of two plant seed ribonucleases from Momordica charantia and Luffa cylindrica" Biosci.Biotech.Biochem.57. (1993)

[文献書誌] Hiroyuki Imai: "Acyl-(acyl-carrier protein)hydrolase from squash cotyledons specific to long-chain fatty acids:purification and characterization" Plant Mol.Biol.20. 199-206 (1992)

[文献書誌] Ikuo Nishida: "The gene and the RNA for the precursor to the plastidlocated glycerol-3-phosphate acyltransferase of Arabidopsis thaliana" Plant Mol.Biol.21. 262-272 (1993)

[文献書誌] Kaoru Suzuki: "Effects of 2,6-dichlorobenzonitrile on differentiation to tracheary elements of isolated mesophylle cells of Zinnia elegans and formation of secondary cell walls" Physiol.Plant. 86. 43-48 (1992)

[文献書誌] Masashi Mori: "Infectivity of plasmids containing brome mosaic virus cDNA linked to the cauliflower mosaic virus 35S RNA promoter" j.Gen.Virology. 72. 243-246 (1991)

[文献書誌] Masahi Mori: "Expression of brome mosaic virus-encoded replicase genes in transgenic tobacco plants" J.Gen.Virology. 73. 169-172 (1992)

[文献書誌] Tsutomu Furuya: "Biotransformation of Tabersonine in Cell Suspension Cultures of Catharunthus roseus" Phytochemistry. 31. 3065-3069 (1992)

[文献書誌] Takafumi Yoshikawa: "Continuous Production of Glycosides by Bioreactor Using Ginseng Hairy Root Cultures" Microbiology and Biothechnology. 38. (1993)

[文献書誌] Koji Mikami: "Regulation of cell cycle-dependent gene expression.In"Control of Plant Gene Expression"(D.P.S.Verma,ed.)" CRC Press,Inc., 18 (1993)

[文献書誌] Takuya Nakayama: "Regulation of wheat histone gene expression.In"Critical Reviews in Plant Science"(B.V.Conger,ed.)" CRC Press,Inc., (1993)

[文献書誌] Hirofumi Uchimiya: "Production of transgenic rice plants by direct gene transfer into protoplasts.In Plant Transformation protocols(ed.L.Fitzmaurice and F.Lin)" Academic Press, (1992)

[文献書誌] Yutaka Oono: "Molecular and cellular analysis of the rol genes of Agrobacterium rhizogenes Ri Plasmid." J.of Plant Research, (1993)

[文献書誌] 米田 好文: "「植物の分子生物学」シロイヌナズナの核ゲノム構成" 共立出版, 372 (1992)

[文献書誌] 米田 好文: "「開花・結実の分子機構」花芽形成開始機構の分子遺伝学的研究" 秀潤社, 290 (1992)

[文献書誌] 中村 研三: "「植物の分子生物学」糖による植物遺伝子の発現制御" 共立出版, 8 (1992)

[文献書誌] 中村 研三: "植物におけるショ糖による遺伝子発現制御" 生化学, 10 (1992)

[文献書誌] 谷口 研至: "苗条原基誘導法「最新バイオテクノロジー全書2 野菜の組織・細胞培養と増殖」" 秀潤社, 8 (1990)

[文献書誌] 谷口 研至: "生長点培養 現代化学増刊「植物バイオテクノジーII」" 東京化学同人, 11 (1991)

[文献書誌] Ikuo Nishida: "Current Topics in Plant Physiology Series" American Society of Plant Physiology, (1993)

[文献書誌] 古谷 力: "Maruzen Advanced Technology 植物工学" 丸善株式会社, 260 (1993)

1992 年度実績報告書

町田泰則名古屋大学, 理学部, 教授 (80175596)

[文献書誌] 渡辺昭: "緑葉の老化とグルタミン合成酵素" Nippon Nogeikagaku Kaishi. 66. 932-935 (1992)

[文献書誌] 米田好文: "「植物の分子生物学」シロイヌナズナの核ゲノム構成" 共立出版, 372 (1992)

[文献書誌] 米田好文: "「開花・結実の分子機構」花芽形成開始機構の分子遺伝学的研究" 秀潤社, 290 (1992)

[文献書誌] 中村研三: "「植物の分子生物学」糖による植物遺伝子の発現制御" 共立出版, 8 (1992)

[文献書誌] 中村研三: "植物におけるショ糖による遺伝子発現制御" 生化学, 10 (1992)

[文献書誌] 谷口研至: "苗条原基誘導法「最新バイオテクノロジー全書2 野菜の組織・細胞培養と増殖」" 秀潤社, 8 (1990)

[文献書誌] 谷口研至: "生長点培養現代化学増刊「植物バイオテクノジーII」" 東京化学同人, 11 (1991)

[文献書誌] 古谷力: "Maruzen Advanced Technology 植物工学" 丸善株式会社, 260 (1993)