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Development of novel gene vectors for the analysis of gene function of non-transformable

Research Project

Project/Area Number 14360205
Research Category

Grant-in-Aid for Scientific Research (B)

Allocation TypeSingle-year Grants
Section一般
Research Field Applied molecular and cellular biology
Research InstitutionYokohama National University

Principal Investigator

HIRATSUKA Kazuyuki  Yokohama National University, Faculty of Environment and Information Sciences, Professor, 大学院・環境情報研究院, 教授 (30202279)

Co-Investigator(Kenkyū-buntansha) NAKAMURA Tatsuo  Yokohama National University, Faculty of Environment and Information Sciences, Lecturer, 大学院・環境情報研究院, 講師 (50334636)
SUZUKI Masashi  Yokohama National University, Faculty of Environment and Information Sciences, Associate Professor, 大学院・環境情報研究院, 助教授 (40282694)
Project Period (FY) 2002 – 2003
Project Status Completed (Fiscal Year 2003)
Budget Amount *help
¥11,500,000 (Direct Cost: ¥11,500,000)
Fiscal Year 2003: ¥3,600,000 (Direct Cost: ¥3,600,000)
Fiscal Year 2002: ¥7,900,000 (Direct Cost: ¥7,900,000)
Keywordsreproduction / meiosis / large-scale analysis / nuclear localization signal / regulated gene expression / transgenic plants / trumpet lily / Arabidopsis / アンチセンス法
Research Abstract

In order to study gene functions in plant cells we developed particle gene delivery system for foreign gene expression in various plant cells. We chose to use Lilium longiflorum as model system because no reliable transformation method is available for lilies. Using lily cDNA library generated from male reproductive organs at various developmental stages, we examined the functional genes involved in microsporogenesis by large-scale sequencing and obtained more than 1000 ESTs. Based on the results obtained by the reverse-northern analysis, we selected number of cDNA clones for further analysis. A cDNA clone, designated M532 showed no extensive sequence homology to previously characterized gene products and specifically expressed during microsporogenesis. Interestingly, the M532 protein targeted to the nucleolus when fused to GFP. Another cDNA clone encoding a protein, designated LISCL, exhibited high similar to the GRAS family. A series of transcriptional activation experiments of truncated LISCL proteins fused to the yeast GAL4 DNA-binding domain clearly demonstrated that the amino terminus of the LISCL protein has a transcriptional activity of the meiosis-associated promoter revealed that in pollen mother cells of lily, the activity of the meiosis-associated promoter is specifically enhanced by the LISCL protin co-expression. These results suggest that the LISCL is involved in the transcriptional regulation during microsporogenesis within the lily anther. We also isolated viral cDNAs from reproductive organs and obtained a full-length cDNA. Sequence analysis revealed that the virus shows strong similarity to the lily symptomless virus. Currently, we are in the process of developing a gene vector system using the full-length infectious clone of the viral cDNA.

Report

(3 results)
  • 2003 Annual Research Report   Final Research Report Summary
  • 2002 Annual Research Report
  • Research Products

    (22 results)

All Other

All Publications (22 results)

  • [Publications] Maeda, T., Watakabe, Y., Seo, S., Takase, H., Hiratsuka, K.: "Expression of the AtRAD51 gene promoter in response to DNA damage in trans genic tobacco."Plant Biotechnol.. 21(in press). (2004)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] Ono, S., Tanaka, T., Watakabe, Y., Hiratsuka, K.: "Transient assay system for the analysis of PR-1a gene promoter in tobacco BY-2 cells."Biosci.Biotechnol.Biochem.. 68(in press). (2004)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] Morohashi, K., Minami, M., Takase, H., Hotta, Y., Hiratsuka, K.: "Isolation and characterization of a novel GRAS gene that regulates meiosis-associated gene expression."J.Biol.Chem.. 278. 20865-20873 (2003)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] Bayarmaa, G., Morohashi, K., Takase, H., Hiratsuka, K.: "Identification of novel microsporogenesis-associated genes encoding proteins with a nuclear localization signal."Plant Biotechnol.. 20. 137-143 (2003)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] George, S.et al.: "Dmc1 fluorescent foci in prophase I nuclei of diploid, triploid and hybrid lilies."Chromosoma. 111. 96-105 (2002)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] Murata, J.et al.: "Characterization of a novel GT-box binding protein from Arabidopsis."Plant Biotechnology. 19. 103-112 (2002)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] 平塚和之: "遺伝子組換え植物の光と影II,導入遺伝子産物による目的以外の影響"学会出版センター. 69-84 (2003)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] Maeda, T., Watakabe, Y., Seo, S., Takase, H., Hiratsuka, K.: "Expression of the AtRAD51 gene promoter in response to DNA damage in transgenic tobacco."Pant Biotechnol.. 21(in press). (2004)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] Ono, S., Tanaka, T., Watakabe, T., Hiratsuka, K.: "Transient assay system for the analysis of PR-1αgene promoter in tobacco BY-2 cells."Biosci.Biotechnol.Biochem.. (in press). (2004)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] Morohashi, K., Minami, M., Takase, H., Hotta, Y., Hiratsuka, K.: "Isolation and characterization of a novel GRAS gene hat regulates meiosis-associated gene expression."J.Biol.Chem.. 278. 20865-20873 (2004)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] Bayarmaa, G., Morohashi, K., Takase, H., Hiratsuka, K.: "Identification of novel microsporogenesis-associated genes encoding proteins with a nuclear localization signal."Plant Biotechniol.. 20. 137-143 (2003)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] Morita, R., Hattori, Y., Yokoi, S., Takase, H., Minami, M., Hiratsuka, K., Toriyama, K.: "Assessment of utility of meiosis-associated promoters of lily for induction of germinal ds transposition in transgenic rice."Plant Cell Physiol.. 44. 637-642 (2003)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] George, S., Behl, P., deGuzman, R., Lee, M., Rusyniak, S., Hotta, Y., Hiratsuka, K., Takase, H., Hasenkampf, C.: Chromosoma. 111. 96-105 (2002)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] Maeda, T., Watakabe, Y., Seo, S., Takase, H., Hiratsuka, K.: "Expression of the AtRAD51 gene promoter in response to DNA damage in transgenic tobacco."Plant Biotechnol.. 21(in press). (2004)

    • Related Report
      2003 Annual Research Report
  • [Publications] Ono, S., Tanaka, T., Watakabe, Y., Hiratsuka, K.: "Transient assay system for the analysis of PR-1a gene promoter in tobacco BY-2 cells."Biosci.Biotechnol.Biochem.. 68(in press). (2004)

    • Related Report
      2003 Annual Research Report
  • [Publications] Morohashi, K., Minami, M., Takase, H., Hotta, Y., Hiratsuka, K.: "Isolation and characterization of a novel GRAS gene that regulates meiosis-associated gene expression."J.Biol.Chem.. 278. 20865-20873 (2003)

    • Related Report
      2003 Annual Research Report
  • [Publications] Bayarmaa, G., Morohashi, K., Takase, H., Hiratsuka, K.: "Identification of novel microsporogenesis-associated genes encoding proteins with a nuclear localization signal."Plant Biotechnol.. 20. 137-143 (2003)

    • Related Report
      2003 Annual Research Report
  • [Publications] Gun-Aajav Bayarmaa, Kengo Morohashi, Hisabumi Takase, Kazuyuki Hiratsuka: "Identification of novel microsporogenesis-associated genes encoding proteins with a nuclear localization signal"Plant Biotechnology. 20(in press). (2003)

    • Related Report
      2002 Annual Research Report
  • [Publications] Morohashi, K., Minami, M., Takase, H., Hotta, Y., Hiratsuka, K.: "Isolation and characterization of a novel GRAS gene that regulates meiosis-associated gene expression"Journal of Biological Chemistry. (in press). (2003)

    • Related Report
      2002 Annual Research Report
  • [Publications] George, S., Behl, P., deGuzman, R., Lee, M., Rusyniak, S., Hotta, Y., Hiratsuka, K., Takase, H., Hasenkampf, C.: "Dmc1 fluorescent foci in prophase I nuclei of diploid, triploid and hydrid lilies"Choromosoma. 111. 96-105 (2002)

    • Related Report
      2002 Annual Research Report
  • [Publications] Murata, J., Takase, H, Hiratsuka, K.: "Characterization of a novel GT-box binding protein from Arabidopsis"Plant Biotechnology. 19. 103-112 (2002)

    • Related Report
      2002 Annual Research Report
  • [Publications] 平塚和之: "次世代の農薬開発"ソフトサイエンス社. 171-180 (2003)

    • Related Report
      2002 Annual Research Report

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Published: 2002-04-01   Modified: 2016-04-21  

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