1990 Fiscal Year Annual Research Report

形質転換と大量集積

Research Project

Project/Area Number	02261103
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, 助手 (10127087) 中村研三名古屋大学, 農学部, 助教授 (80164292) 内宮博文北海道大学, 理学部, 助教授 (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 / シコニン合成
Research Abstract	本研究計画の目標は、(1)植物の生産する有用物質大量集積を担っている遺伝子発現様式を、トランスジェニック植物を用いて解明すること、(2)有用物質を生産している培養系を用いて、その鍵となっている新しい遺伝子をクロ-ン化すること、さらにこれらを統一し最終的に有用物質大量集積の方途を探ることである。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-からのTーDNA転移頻度を決定しているvir遺伝子の発現を、植物細胞壁多糖の構成糖が著しく促進することを見いだした。また、植物染色体へのTーDNAの組み込みに関与していると思われる染色体上の塩基配列を見つけた。(2)植物ヒストン遺伝子の転写因子のHBP1ーa,HBP1ーbの構造を明らかにした。(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、そのcis領域を同定した。(4)酵母細胞中で窒素除去に応答するシロイヌナズナDNA断片を単離し、植物に再導入したところ、植物中でも窒素除去に応答することがわかった。(5)ショ糖で発現が誘導される貯蔵タンパク質スポラミン遺伝子のプロモ-タ-のcis領域を決定した。スポラミンと同様,塊根で発現しているβーアミラ-ゼ遺伝子もショ糖で誘導されること、さらにこれら二つの遺伝子のプロモ-タ-領域に共通に結合するタンパク質因子を見つけた。(6)ベルベリン生合成の律速酵素段階を特定するとともに二つのメチルトランスフェラ-ゼを単一に精製した。(7)培養細胞系を用いてシコニン生合成の内在性誘導物質、及びベルベリンの生成と分泌に要する時間等を解明した。(8)苗条原基からリピッドボディの単離を可能にし、また苗条原基に遺伝子を導入する方法を検討した。(9)薬用植物への遺伝子導入系を確率し、実際にマウスのチトクロ-ムPー450遺伝子を導入した植物を作製した。(10)ブドウ培養細胞のケルセチン配糖化酵素の精製を行い、その細胞内局在部位を調べた。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

(19 results)

All Other

All Publications (19 results)

[Publications] Nobuyoshi Shimoda: "Control of expression of <Agrobacterium>___ー vir genes by synergistic actions of phenolic signal molecules and monosaccharides" Proceedings of National Academy of Science,USA. 87. 6684-6688 (1990)
[Publications] Shogo Matsumoto: "Integration of <Agrobacterium>___ー TーDNA into a tobacco chromosome:Possible involvement of DNA homology between TーDNA and plant DNA" Molecular General Genetics. 224. 309-316 (1990)
[Publications] Takefumi Kawata: "Cell cycleーregulated gene expression in transgenic plant cells" Develop.Genet.11. 205-213 (1990)
[Publications] Koji Mikami: "Regulation of cell cycleーdependent gene expression.In“Control of Plant Gene Expression"" Telford Press. (1990)
[Publications] Kiyoshi Kanaya: "Specific binding of nuclear protein from tobacco hairy roots cultured in Vitro to a 5'ーupstream region of the rolC gene of the Ri plasmid" Plant Cell Physiol.31. 941-946 (1990)
[Publications] Nobuharu Fujii: "Conditions favorable for the somatic embryogenesis in carrot cell culture enhance expression of the <rolC>___ー promoterーGUS fusion gene" Plant Physiol.95. 238-241 (1991)
[Publications] Takashi Araki: "Electrophoretic analysis of florallyーevoked meristems of <Pharbitis>___ー <nil>___ー Choisy cv.violet." Plant Cell Physiol.31. 137-144 (1990)
[Publications] Tsukaho Hattori: "Sucroseーinduced expression of genes coding for tuberous root storage protein,sporamin,of sweet potato in leave and petioles" Plant & Cell Physiol.32. 79-86 (1991)
[Publications] Shozo Ohta: "Stemーpredominant expression of the sweet potato sporamin gene promoter:βーglucuronidase(GUS)furion gene in transgenic tobacco is conferred by multiple cell typeーspecfic regulatory elements" Mol.Gen.Genet.
[Publications] Fumiko,Sato: "Synthesis of plant triosephosphate isomerase in <Escherichia coli>___ー." Agric.Biol.chem.54. 2189-2191 (1990)
[Publications] Hiroshi Fukui: "Induction of shikonin biosynthesis by endogenous polysaccharides in <Lithospermum>___ー <erythrorhizon>___ー cell sespension cultures" Plant Cell Reports. 9. 73-76 (1990)
[Publications] Hideharu Sato: "Specific differences in tolerance to exogenous berberine among plant cell cultures" Plant Cell Reports. 9. 133-136 (1990)
[Publications] Toru Kodama: "Glucoylation of Quercetin by Cell Suspension Culture of Vitis sp." Agric.Biol.Chem.54. 3283-3288 (1990)
[Publications] Tetsuro Kokubo: "Quercetin 3ーOーβーDーGlucopyranoside and Isorhamnetin 3ーOーβーDーGlucopyranoside Formation from Quercetin by Cell Culdtures of Ipomoea batatas and Crocus sativum." Agric.Biol.Chem.55. 613-614 (1991)
[Publications] 谷口研至: "苗条原基誘導法" 最新バイオテクノロジ-全書2・野菜の組織・細胞培養と増殖. 280-287 (1990)
[Publications] 谷口研至: "苗条原基" 植物細胞工学. 2. 249-255 (1990)
[Publications] Kazuki Saito: "Stable transfer and expression of chimeric genes in licorice(<Glycyrrhiza uralensis>___ー)using an Ri plasmid binary vector" Plant Cell Reports. 8. 718-721 (1990)
[Publications] Kazuki Saito: "Genetic transformation of foxglove(<Digitalis purpurea>___ー)by chimeric foreign genes and production of cardioactive glycosides" Plant Cell Reports. 9. 121-124 (1990)
[Publications] 内宮博文: "植物遺伝子操作マニュアルートランスジェニック植物の作り方ー" 講談社, 138 (1990)

1990 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] Nobuyoshi Shimoda: "Control of expression of <Agrobacterium>___ー vir genes by synergistic actions of phenolic signal molecules and monosaccharides" Proceedings of National Academy of Science,USA. 87. 6684-6688 (1990)

[Publications] Shogo Matsumoto: "Integration of <Agrobacterium>___ー TーDNA into a tobacco chromosome:Possible involvement of DNA homology between TーDNA and plant DNA" Molecular General Genetics. 224. 309-316 (1990)

[Publications] Takefumi Kawata: "Cell cycleーregulated gene expression in transgenic plant cells" Develop.Genet.11. 205-213 (1990)

[Publications] Koji Mikami: "Regulation of cell cycleーdependent gene expression.In“Control of Plant Gene Expression"" Telford Press. (1990)

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

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

[Publications] Takashi Araki: "Electrophoretic analysis of florallyーevoked meristems of <Pharbitis>_ー <nil>_ー Choisy cv.violet." Plant Cell Physiol.31. 137-144 (1990)

[Publications] Tsukaho Hattori: "Sucroseーinduced expression of genes coding for tuberous root storage protein,sporamin,of sweet potato in leave and petioles" Plant & Cell Physiol.32. 79-86 (1991)

[Publications] Shozo Ohta: "Stemーpredominant expression of the sweet potato sporamin gene promoter:βーglucuronidase(GUS)furion gene in transgenic tobacco is conferred by multiple cell typeーspecfic regulatory elements" Mol.Gen.Genet.

[Publications] Fumiko,Sato: "Synthesis of plant triosephosphate isomerase in <Escherichia coli>___ー." Agric.Biol.chem.54. 2189-2191 (1990)

[Publications] Hiroshi Fukui: "Induction of shikonin biosynthesis by endogenous polysaccharides in <Lithospermum>_ー <erythrorhizon>_ー cell sespension cultures" Plant Cell Reports. 9. 73-76 (1990)

[Publications] Hideharu Sato: "Specific differences in tolerance to exogenous berberine among plant cell cultures" Plant Cell Reports. 9. 133-136 (1990)

[Publications] Toru Kodama: "Glucoylation of Quercetin by Cell Suspension Culture of Vitis sp." Agric.Biol.Chem.54. 3283-3288 (1990)

[Publications] Tetsuro Kokubo: "Quercetin 3ーOーβーDーGlucopyranoside and Isorhamnetin 3ーOーβーDーGlucopyranoside Formation from Quercetin by Cell Culdtures of Ipomoea batatas and Crocus sativum." Agric.Biol.Chem.55. 613-614 (1991)

[Publications] 谷口研至: "苗条原基誘導法" 最新バイオテクノロジ-全書2・野菜の組織・細胞培養と増殖. 280-287 (1990)

[Publications] 谷口研至: "苗条原基" 植物細胞工学. 2. 249-255 (1990)

[Publications] Kazuki Saito: "Stable transfer and expression of chimeric genes in licorice(<Glycyrrhiza uralensis>___ー)using an Ri plasmid binary vector" Plant Cell Reports. 8. 718-721 (1990)

[Publications] Kazuki Saito: "Genetic transformation of foxglove(<Digitalis purpurea>___ー)by chimeric foreign genes and production of cardioactive glycosides" Plant Cell Reports. 9. 121-124 (1990)

[Publications] 内宮博文: "植物遺伝子操作マニュアルートランスジェニック植物の作り方ー" 講談社, 138 (1990)

1990 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] Nobuyoshi Shimoda: "Control of expression of <Agrobacterium>___ー vir genes by synergistic actions of phenolic signal molecules and monosaccharides" Proceedings of National Academy of Science,USA. 87. 6684-6688 (1990)

[Publications] Shogo Matsumoto: "Integration of <Agrobacterium>___ー TーDNA into a tobacco chromosome:Possible involvement of DNA homology between TーDNA and plant DNA" Molecular General Genetics. 224. 309-316 (1990)

[Publications] Takefumi Kawata: "Cell cycleーregulated gene expression in transgenic plant cells" Develop.Genet.11. 205-213 (1990)

[Publications] Koji Mikami: "Regulation of cell cycleーdependent gene expression.In“Control of Plant Gene Expression"" Telford Press. (1990)

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

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

[Publications] Takashi Araki: "Electrophoretic analysis of florallyーevoked meristems of <Pharbitis>___ー <nil>___ー Choisy cv.violet." Plant Cell Physiol.31. 137-144 (1990)

[Publications] Tsukaho Hattori: "Sucroseーinduced expression of genes coding for tuberous root storage protein,sporamin,of sweet potato in leave and petioles" Plant & Cell Physiol.32. 79-86 (1991)

[Publications] Shozo Ohta: "Stemーpredominant expression of the sweet potato sporamin gene promoter:βーglucuronidase(GUS)furion gene in transgenic tobacco is conferred by multiple cell typeーspecfic regulatory elements" Mol.Gen.Genet.

[Publications] Fumiko,Sato: "Synthesis of plant triosephosphate isomerase in <Escherichia coli>___ー." Agric.Biol.chem.54. 2189-2191 (1990)

[Publications] Hiroshi Fukui: "Induction of shikonin biosynthesis by endogenous polysaccharides in <Lithospermum>___ー <erythrorhizon>___ー cell sespension cultures" Plant Cell Reports. 9. 73-76 (1990)

[Publications] Hideharu Sato: "Specific differences in tolerance to exogenous berberine among plant cell cultures" Plant Cell Reports. 9. 133-136 (1990)

[Publications] Toru Kodama: "Glucoylation of Quercetin by Cell Suspension Culture of Vitis sp." Agric.Biol.Chem.54. 3283-3288 (1990)

[Publications] Tetsuro Kokubo: "Quercetin 3ーOーβーDーGlucopyranoside and Isorhamnetin 3ーOーβーDーGlucopyranoside Formation from Quercetin by Cell Culdtures of Ipomoea batatas and Crocus sativum." Agric.Biol.Chem.55. 613-614 (1991)

[Publications] 谷口 研至: "苗条原基誘導法" 最新バイオテクノロジ-全書2・野菜の組織・細胞培養と増殖. 280-287 (1990)

[Publications] 谷口 研至: "苗条原基" 植物細胞工学. 2. 249-255 (1990)

[Publications] Kazuki Saito: "Stable transfer and expression of chimeric genes in licorice(<Glycyrrhiza uralensis>___ー)using an Ri plasmid binary vector" Plant Cell Reports. 8. 718-721 (1990)

[Publications] Kazuki Saito: "Genetic transformation of foxglove(<Digitalis purpurea>___ー)by chimeric foreign genes and production of cardioactive glycosides" Plant Cell Reports. 9. 121-124 (1990)

[Publications] 内宮 博文: "植物遺伝子操作マニュアルートランスジェニック植物の作り方ー" 講談社, 138 (1990)

町田泰則名古屋大学, 理学部, 教授 (80175596)

[Publications] Takashi Araki: "Electrophoretic analysis of florallyーevoked meristems of <Pharbitis>_ー <nil>_ー Choisy cv.violet." Plant Cell Physiol.31. 137-144 (1990)

[Publications] Hiroshi Fukui: "Induction of shikonin biosynthesis by endogenous polysaccharides in <Lithospermum>_ー <erythrorhizon>_ー cell sespension cultures" Plant Cell Reports. 9. 73-76 (1990)

[Publications] 谷口研至: "苗条原基誘導法" 最新バイオテクノロジ-全書2・野菜の組織・細胞培養と増殖. 280-287 (1990)

[Publications] 谷口研至: "苗条原基" 植物細胞工学. 2. 249-255 (1990)

[Publications] 内宮博文: "植物遺伝子操作マニュアルートランスジェニック植物の作り方ー" 講談社, 138 (1990)