• Search Research Projects
  • Search Researchers
  • How to Use
  1. Back to previous page

cDNAセレクション法とトランスポゾンタギング法による高等植物の機能遺伝子の単離

Research Project

Project/Area Number 09780647
Research Category

Grant-in-Aid for Encouragement of Young Scientists (A)

Allocation TypeSingle-year Grants
Research Field Molecular biology
Research InstitutionThe Institute of Physical and Chemical Research

Principal Investigator

関 原明  理化学研究所, 植物分子生物学研究室, 研究員 (80281624)

Project Period (FY) 1997 – 1998
Project Status Completed (Fiscal Year 1998)
Budget Amount *help
¥2,000,000 (Direct Cost: ¥2,000,000)
Fiscal Year 1998: ¥900,000 (Direct Cost: ¥900,000)
Fiscal Year 1997: ¥1,100,000 (Direct Cost: ¥1,100,000)
Keywordsシロイヌナズナ / トランスポゾンタギング / Ac / Ds / ラテックス粒子 / cDNAセレクション法 / RD28遺伝子 / Receptor-like protein kinase
Research Abstract

cDNAスキャンニング法とトランスポゾンタギング法を組み合わせて単離した植物遺伝子の機能を解析する方法の開発に着手した。シロイヌナズナの5番染色体にDsT-DNAが挿入された親植物系統(Ds388-30)を用いてDsを転移させ、これまでに転移植物系統(1000系統)を作製した。Dsドナー部位を含む2つのYACクローン(CIC5Fll,CIC2B9)に対してcDNAスキャンニング法によりcDNAを単離した。PCRスクリーニングの結果、単離された遺伝子のreceptor-like protein kinaseのホモログ遺伝子およびシロイヌナズナのESTcDNA(T42369)と相同な遺伝子の遺伝子内部にDsが挿入した系統が選抜された。
また、転移系統(200系統)よりTAIL-PCR法を用いてDsの挿入により破壊された遺伝子断片を回収し塩基配列を決定した。その結果、endoxyloglucan transferase遺伝子(EXGT-A3),actin-4遺伝子,water channel protein遺伝子(RD28)、オーキシンの抵抗性に関与するAUX1遺伝子のホモログ遺伝子,WDrepeat protein ATANllのホモログ遺伝子,PP2Aのβ'調節サブユニット遺伝子などの遺伝子内部にDsが挿入した系統が作製されていることがわかった。現在、これらの系統についてDsの挿入変異をホモにもつ個体の表現型を調べることにより破壊された遺伝子の機能解析を進めている。

Report

(2 results)
  • 1998 Annual Research Report
  • 1997 Annual Research Report
  • Research Products

    (14 results)

All Other

All Publications (14 results)

  • [Publications] Seki, M.: "High-efficiency cloning of Arabidopsis fall-length cDNA by biotinylated CAP trapper" Plant Journal. 15(5). 707-720 (1998)

    • Related Report
      1998 Annual Research Report
  • [Publications] Shinozaki, K.: "Molecular Responses to Water Stress in Arabidopsis thaliana" Journal of Plant Research. 111. 345-351 (1998)

    • Related Report
      1998 Annual Research Report
  • [Publications] Shinwari, Z.K.: "An Arabidopsis Gene Family Encoding DRE/CRT Binding Protein Involved in Low-Temperature-Responsive Gene Expression" Biochemical and Biophysical Research Communications. 250. 161-170 (1998)

    • Related Report
      1998 Annual Research Report
  • [Publications] Ichimura, K.: "Molecular Cloning and Characterization of Three cDNAs Encoding Putative Mitogen-activated Protein Kinase Kinases(MAPKKs)in Arabidopsis thaliana" DNA Research. 5. 341-348 (1998)

    • Related Report
      1998 Annual Research Report
  • [Publications] Shibata, D.: "Establishment of framework PI clones for map-based cloning and genome sequencing:direct RFLP mapping of large clones" Gene. 225. 31-38 (1998)

    • Related Report
      1998 Annual Research Report
  • [Publications] Taji, T.: "Mapping of 25 Drought-Inducible Genes, RD and ERD, in Arabidopsis thaliana" Plant and Cell Physiology. 40(1). 119-123 (1999)

    • Related Report
      1998 Annual Research Report
  • [Publications] Ito, T.: "Regional insertional mutagenesis of genes on Arabidopsis thaliana chromosome V using the Ac/Ds transposon in combination with a cDNA scanning method" Plant Journal. (in press). (1999)

    • Related Report
      1998 Annual Research Report
  • [Publications] Seki, M.: "Transient expression of foreign genes in tissues of Arabidopsis thaliana by bombardment-mediated transformation. Methods in Molecular Biology. Vol.82" Eds.by Salinas, J.and Martinez-Zapater, J.M.HUMANA PRESS INC., 7 (1998)

    • Related Report
      1998 Annual Research Report
  • [Publications] 伊藤卓也: "遺伝子ライブラリーの調製.新版微生物学実験法.大和田紘一ら(編)(in press)" 講談社 サイエンティフィク, (1999)

    • Related Report
      1998 Annual Research Report
  • [Publications] Seki, M.: "Transient expression of foreign genes in tissues of Arabidopsis thaliana by bombardment-mediated transformation. Molecular Biotechnology(in press)" Ed.by Walker, J.M.HUMANA PRESS INC., (1999)

    • Related Report
      1998 Annual Research Report
  • [Publications] Seki,M.: "Rapid construction of a franscriptoin map for a cosmid contig of Arabidopsis thaliana genome using a novel cDNA selection method" Plant Journal. 12(2). 481-487 (1997)

    • Related Report
      1997 Annual Research Report
  • [Publications] Inada,H.: "Existence of three regulatory regions each,containing a highly conserved motif in the promoter of plastid-encoded RNA polymerase gene (rpoB)" Plant Journal. 11(4). 883-890 (1997)

    • Related Report
      1997 Annual Research Report
  • [Publications] Seki,M.: "Transient expression of foreign genes in tissues of Arabidopsis thaliana by bombardment mediated transformation. Methods in Molecular Biology." Eds.by Slinas,J.and Martinez-Zapater,J.M.HUMANA PRESS INC., 7 (1997)

    • Related Report
      1997 Annual Research Report
  • [Publications] 伊藤卓也: "遺伝子ライブラリーの調製、新版、微生物学実験法 大和田紘一ら(編)" 講談社サイエンティフィク(発表予定),

    • Related Report
      1997 Annual Research Report

URL: 

Published: 1997-04-01   Modified: 2016-04-21  

Information User Guide FAQ News Terms of Use Attribution of KAKENHI

Powered by NII kakenhi