1991 Fiscal Year Annual Research Report

形質転換と大量集積

Research Project

Project/Area Number	03257103
Research Institution	Nagoya University
Principal Investigator	町田泰則名古屋大学, 理学部, 教授 (80175596)
Co-Investigator(Kenkyū-buntansha)	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, 助教授 (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)
Keywords	Tiプラスミドベクタ- / 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-ゼ
Research Abstract	本年度は、以下のような成果を挙げた。 (1)Tiプラスミドベクタ-による形質転換機構を研究して、その頻度が上昇する変異株を分離し、さらに、その変異遺伝子タンパク質と相互作用するタンパク質を見いだした。一方、シンク器官の一つである種子で発現する遺伝子プロモ-タ-を解析した所、丁でも高発現していることがわかり、これを根での人工的な物質生産系に応用できる可能性を示した。(2)植物ヒストン遺伝子の転写因子HBP1ーaの機能解析の結果、これと相互作用するDNA非結合型転写制御因子の存在が示唆された。(3)物質の輸送に関与している師部組織特異的に発現しているrolC遺伝子のプロモ-タ-を解析し、0.4モルショ糖により活性化されることがわかった。(4)シロイヌナズナの窒素除去に応答する遺伝子プロモ-タ-を解析し、必須領域を600bp以内に限定した。(5)ショ糖で発現が誘導されるスポラミン遺伝子とβーアミラ-ゼのプロモ-タ-には、共通に必要とされる配列があり、その内の一つに結合するタンパク質をコ-ドしているcDNAを単離して構造を決定した。(6)ベルベリン生合成酵素の精製ならびにその酵素化学的性質の解析、さらに部分アミノ酸配列の決定を行った。(7)培養細胞におけるベルベリン、シコニン、及びブリオノ-ル酸の生合成調節因子と集積動態を明らかにした。(8)カミツレ苗条原基から油体を単離し、構成成分を調べたところ、主としてアシルグリセロ-ルと遊離脂肪酸及び21kDaの特異的タンパク質が認められた。(9)高等植物から世界で初めて含硫アミノ酸生合成の鍵酵素であるシステイン合成酵素のcDNAをクロ-ニングした。このクロ-ンはCys^-大腸菌のCys要求性を回復した。(10)エンドウ芽生えよりオキシドスクワレン:シクロアルテノ-ルサイクレ-ルを精製した。(11)老化緑葉に発現するグルタミン合成酵素遺伝子を単離し、同酵素を持たないタバコに導入した。(12)ヘチマ種子主要タンパク質を調べ、その内luffinはそのC末端の10アミノ酸が除去されると細胞外に集積されることを明らかにした。(13)植物脂肪酸代謝の鍵酵素であるアシルAcpヒドロラ-ゼをカボチャより精製し、部分一次構造を決定した。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。

Research Products

(28 results)

All Other

All Publications (28 results)

[Publications] Shigehisa Okamoto: "Localization and orientation of the VirD4 protein of Agrobacterium tumefaciens in the cell membrane" Molecular General Genetics. 228. 24-32 (1991)
[Publications] Hitoshi Onouchi: "Operation of an efficient site-specific recombination system of Zygosaccharomyces rouxii in togacco cells" Nucleic Acids Research. 19. 6373-6378 (1991)
[Publications] Tetsuya Tabata: "HBP-1a and HBP-1b:leucine zipper-type transcription factors of wheat" EMBO J.10. 1459-1467 (1991)
[Publications] Hisabumi Takase: "Sequence-specific single strand DNA-binding proteins that interact with the regulatory regions of wheat histone H3 and H4 genes" Biochem.Biophys.Res.Commun.176. 1593-1600 (1991)
[Publications] Kiyoshi Kanaya: "Specific binding of nuclear protein from tobacco hairy roots cultured in virto to a 5'-upstream region of the colC gene of the Ri plasmid" Plant Cell Physiology. 31. 941-946 (1990)
[Publications] Nobuharu Fujii: "Conditions favorable for the somatic embryogenisis in carrot cell culture enhance expression of the rolC promoter-GUS fusion gene" Plant Physiology. 95. 238-241 (1991)
[Publications] Kenzo Nakamura: "Sucrose-induced accumulation of β-amylase occurs concomitant with the accumulation of starch and sporamin in leaf-petiole cuttings of the sweet potato" Plant Physiology. 96. 902-909 (1991)
[Publications] Sumiei Ishiguro: "The nuclear factor SP8BF binds to the 5'-upstream regions of three different genes conding for major tuberous root proteins of sweet potato" Plant Molecular Biology. 18. 97-108 (1992)
[Publications] Yoshitaka Azumi: "Evidence for a senescence-associated geneinduced by darkness" Plant Physiol.95. 577-583 (1991)
[Publications] Yoshitaka Azumi: "Structure and regulation by suger of dinl,a dark-inducible and senescence-associated gene in radish"
[Publications] Nozomu Koizumi: "Sequence analysis of cDNA encoding phosphoenolpyruvate carboxylase from cultured tobacco cells" Plant Molecular Biology. 17. 535-539 (1991)
[Publications] Satomi Takeda: "Nucleotide sequence of a cDNA for Osmotin-like protein from cultured tobacco cells" Plant Physiol.97. 844-846 (1991)
[Publications] Mamoru Tabata: "Transport and secretion of natural products in plant cell cultures" Planta Medica. 57. 21-26 (1991)
[Publications] Yoshinori Kobayashi: "The role of ethylene in berberine production by Thalictrum minus cell suspension cultures" Phytochemistry. 30. 3605-3609 (1991)
[Publications] Song Yang-Nong: "Synergistic action of Phenolic Signal Compounds and Carbohydrates in the Induction of Virlence Gene Expression of Agrobacterium tumefaciens" Chem.Pharm.Bull.39. 2613-2616 (1991)
[Publications] Ikuro Abe: "Purification of 2.3-Oxido-spualene:Cycloartenol Cyclase and:β-Amyrin Cyclase from Pea Seedlings" Chem.Pharm.Bull.40. (1992)
[Publications] Hiroyuki Takano: "Rapid clonal-propagation of Matricaria chamomilla by tissuecultured shoot primordia" Japanese Journal of Genetics. 41. 421-426 (1991)
[Publications] Kenji Taniguchi: "Visualization of replicating bands in plant chromosomes with a monoclonal anti-BrdU antibody method" Japanese Journal of Genetics. 66. 485-489 (1991)
[Publications] Kazuki Saito: "Integration and expression of a rabbit liver cytochrome P-450 gene in transgenic Nicotiana tabacum" The Proceedings of The National Academy of Science USA. 88. 7041-7045 (1991)
[Publications] Kazuki Saito: "Tissue-specific and stress-enhancing expression of the TR promoter for mannopine synthase in transgenic plants" Planta. 184. 40-46 (1991)
[Publications] Tomomitsu Hatayama: "Amino acid sequences of the two smallest trypsin inhibitors from sponge gourd seeds" Agric.Biol.Chem.55. 2641-2642 (1991)
[Publications] Gunki Funatsu: "Conserved amino acid residues in ribosome-inactivating proteins from plasts" Biochemie. 73. 1157-1161 (1991)
[Publications] Hiroyuki Imai: "Acyl-(acyl-carrier-protein) hydorolase from squash cotyledons specific to long-chain fatty acids:Purification and characterization" Plant molecular Biology. (1992)
[Publications] 内宮博文: "植物遺伝子操作マニュアル" 講談社, 138 (1990)
[Publications] 中村研三: "『植物全能性の分子生物学』「植物遺伝子の器官特異的な発現とその柔軟性」" 朝倉書店, 22 (1991)
[Publications] 中村研三: "『植物バイオテクノロジ-II』「サツマイモのスポラミン遺伝子の形質転換植物での発現」" 東京化学同人, 10 (1991)
[Publications] 谷口研至: "『植物細胞工学』苗条原基" 秀潤社, 7 (1990)
[Publications] 谷口研至: "『植物バイオテクノロジ-II』生長点培養" 東京化学同人, 11 (1991)

1991 Fiscal Year Annual Research Report

形質転換と大量集積

Principal Investigator

町田 泰則 名古屋大学, 理学部, 教授 (80175596)

Research Products

[Publications] Shigehisa Okamoto: "Localization and orientation of the VirD4 protein of Agrobacterium tumefaciens in the cell membrane" Molecular General Genetics. 228. 24-32 (1991)

[Publications] Hitoshi Onouchi: "Operation of an efficient site-specific recombination system of Zygosaccharomyces rouxii in togacco cells" Nucleic Acids Research. 19. 6373-6378 (1991)

[Publications] Tetsuya Tabata: "HBP-1a and HBP-1b:leucine zipper-type transcription factors of wheat" EMBO J.10. 1459-1467 (1991)

[Publications] Hisabumi Takase: "Sequence-specific single strand DNA-binding proteins that interact with the regulatory regions of wheat histone H3 and H4 genes" Biochem.Biophys.Res.Commun.176. 1593-1600 (1991)

[Publications] Kiyoshi Kanaya: "Specific binding of nuclear protein from tobacco hairy roots cultured in virto to a 5'-upstream region of the colC gene of the Ri plasmid" Plant Cell Physiology. 31. 941-946 (1990)

[Publications] Nobuharu Fujii: "Conditions favorable for the somatic embryogenisis in carrot cell culture enhance expression of the rolC promoter-GUS fusion gene" Plant Physiology. 95. 238-241 (1991)

[Publications] Kenzo Nakamura: "Sucrose-induced accumulation of β-amylase occurs concomitant with the accumulation of starch and sporamin in leaf-petiole cuttings of the sweet potato" Plant Physiology. 96. 902-909 (1991)

[Publications] Sumiei Ishiguro: "The nuclear factor SP8BF binds to the 5'-upstream regions of three different genes conding for major tuberous root proteins of sweet potato" Plant Molecular Biology. 18. 97-108 (1992)

[Publications] Yoshitaka Azumi: "Evidence for a senescence-associated geneinduced by darkness" Plant Physiol.95. 577-583 (1991)

[Publications] Yoshitaka Azumi: "Structure and regulation by suger of dinl,a dark-inducible and senescence-associated gene in radish"

[Publications] Nozomu Koizumi: "Sequence analysis of cDNA encoding phosphoenolpyruvate carboxylase from cultured tobacco cells" Plant Molecular Biology. 17. 535-539 (1991)

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

[Publications] Mamoru Tabata: "Transport and secretion of natural products in plant cell cultures" Planta Medica. 57. 21-26 (1991)

[Publications] Yoshinori Kobayashi: "The role of ethylene in berberine production by Thalictrum minus cell suspension cultures" Phytochemistry. 30. 3605-3609 (1991)

[Publications] Song Yang-Nong: "Synergistic action of Phenolic Signal Compounds and Carbohydrates in the Induction of Virlence Gene Expression of Agrobacterium tumefaciens" Chem.Pharm.Bull.39. 2613-2616 (1991)

[Publications] Ikuro Abe: "Purification of 2.3-Oxido-spualene:Cycloartenol Cyclase and:β-Amyrin Cyclase from Pea Seedlings" Chem.Pharm.Bull.40. (1992)

[Publications] Hiroyuki Takano: "Rapid clonal-propagation of Matricaria chamomilla by tissuecultured shoot primordia" Japanese Journal of Genetics. 41. 421-426 (1991)

[Publications] Kenji Taniguchi: "Visualization of replicating bands in plant chromosomes with a monoclonal anti-BrdU antibody method" Japanese Journal of Genetics. 66. 485-489 (1991)

[Publications] Kazuki Saito: "Integration and expression of a rabbit liver cytochrome P-450 gene in transgenic Nicotiana tabacum" The Proceedings of The National Academy of Science USA. 88. 7041-7045 (1991)

[Publications] Kazuki Saito: "Tissue-specific and stress-enhancing expression of the TR promoter for mannopine synthase in transgenic plants" Planta. 184. 40-46 (1991)

[Publications] Tomomitsu Hatayama: "Amino acid sequences of the two smallest trypsin inhibitors from sponge gourd seeds" Agric.Biol.Chem.55. 2641-2642 (1991)

[Publications] Gunki Funatsu: "Conserved amino acid residues in ribosome-inactivating proteins from plasts" Biochemie. 73. 1157-1161 (1991)

[Publications] Hiroyuki Imai: "Acyl-(acyl-carrier-protein) hydorolase from squash cotyledons specific to long-chain fatty acids:Purification and characterization" Plant molecular Biology. (1992)

[Publications] 内宮 博文: "植物遺伝子操作マニュアル" 講談社, 138 (1990)

[Publications] 中村 研三: "『植物全能性の分子生物学』「植物遺伝子の器官特異的な発現とその柔軟性」" 朝倉書店, 22 (1991)

[Publications] 中村 研三: "『植物バイオテクノロジ-II』「サツマイモのスポラミン遺伝子の形質転換植物での発現」" 東京化学同人, 10 (1991)

[Publications] 谷口 研至: "『植物細胞工学』苗条原基" 秀潤社, 7 (1990)

[Publications] 谷口 研至: "『植物バイオテクノロジ-II』生長点培養" 東京化学同人, 11 (1991)

町田泰則名古屋大学, 理学部, 教授 (80175596)

[Publications] 内宮博文: "植物遺伝子操作マニュアル" 講談社, 138 (1990)

[Publications] 中村研三: "『植物全能性の分子生物学』「植物遺伝子の器官特異的な発現とその柔軟性」" 朝倉書店, 22 (1991)

[Publications] 中村研三: "『植物バイオテクノロジ-II』「サツマイモのスポラミン遺伝子の形質転換植物での発現」" 東京化学同人, 10 (1991)

[Publications] 谷口研至: "『植物細胞工学』苗条原基" 秀潤社, 7 (1990)

[Publications] 谷口研至: "『植物バイオテクノロジ-II』生長点培養" 東京化学同人, 11 (1991)