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Development of caspase-3 indicator based-on bioluminescence-fluorescence double resonant energy transfer

Research Project

Project/Area Number 20770115
Research Category

Grant-in-Aid for Young Scientists (B)

Allocation TypeSingle-year Grants
Research Field Biophysics
Research InstitutionHokkaido University

Principal Investigator

SAITO Kenta  Hokkaido University, 電子科学研究所, 特任准教授 (60374659)

Project Period (FY) 2008 – 2010
Project Status Completed (Fiscal Year 2010)
Budget Amount *help
¥4,550,000 (Direct Cost: ¥3,500,000、Indirect Cost: ¥1,050,000)
Fiscal Year 2010: ¥390,000 (Direct Cost: ¥300,000、Indirect Cost: ¥90,000)
Fiscal Year 2009: ¥910,000 (Direct Cost: ¥700,000、Indirect Cost: ¥210,000)
Fiscal Year 2008: ¥3,250,000 (Direct Cost: ¥2,500,000、Indirect Cost: ¥750,000)
Keywordsバイオイメージング / 細胞内情報伝達 / 生物発光共鳴エネルギー移動 / 機能イメージング / カルシウム指示薬 / BRET / イメージング / Rluc / Venus / 発光タンパク / 蛍光タンパク質 / 自家蛍光
Research Abstract

In this study, first, bioluminescence resonance energy transfer (BRET) based Ca^<2+> indicator, BRAC was developed and tested to evaluate its physiochemical properties. Second, BRAC was used to visualize Ca^<2+> dynamics in living cells (HeLa cells) and living plant (Arabidopsis thaliana) leaf. Finally, to make the indicator based on bioluminescent-fluorescent double resonant energy transfer, appropriate FRET acceptor for FRET donor Venus were investigated in possible orange to red fluorescent protein candidates. However FRET efficiency between Venus and these orange to red fluorescent proteins were poor, so additional studies should be needed.

Report

(4 results)
  • 2010 Annual Research Report   Final Research Report ( PDF )
  • 2009 Annual Research Report
  • 2008 Annual Research Report
  • Research Products

    (9 results)

All 2010 2008 Other

All Journal Article (3 results) (of which Peer Reviewed: 3 results) Presentation (3 results) Remarks (3 results)

  • [Journal Article] Auto-luminescent genetically-encoded ratiometric indicator for real-time Ca2+ imaging at the single cell level.2010

    • Author(s)
      Kenta Saito, Noriyuki Hatsugai, Kazuki Horikawa, Kentaro Kobayashi, Toru Matsu-Ura, Katsuhiko Mikoshiba, Takeharu Nagai.
    • Journal Title

      PLoS ONE 5

    • NAID

      120002063214

    • Related Report
      2010 Final Research Report
    • Peer Reviewed
  • [Journal Article] Auto-luminescent genetically-encoded ratiometric indicator for real-time Ca2+ imaging at the single cell level.2010

    • Author(s)
      Kenta Saito, Noriyuki Hatsugai, Kazuki Horikawa, Kentaro Kobayashi, Toru Matsu-ura, Katsuhiko Mikoshiba, Takeharu Nagai
    • Journal Title

      PLoS ONE

      Volume: 5

    • NAID

      120002063214

    • Related Report
      2010 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Auto-luminescent genetically-encoded ratiometric indicator for real-time Ca2+ imaging at the single cell level2010

    • Author(s)
      Kenta Saito, Noriyuki Hatsugai, Kazuki Horikawa, Kentaro Kobayashi, Toru Matsu-ura, Katsuhiko Mikoshiba, Takeharu Nagai
    • Journal Title

      PloS ONE 5

    • NAID

      120002063214

    • Related Report
      2009 Annual Research Report
    • Peer Reviewed
  • [Presentation] Auto-luminescent genetically-encoded ratiometric indicator for real-time Ca2+ imaging at the single cell level.2010

    • Author(s)
      Kenta Saito, Takeharu Nagai
    • Organizer
      ECS 2010 Warsaw
    • Place of Presentation
      Warsaw Ochota Campus・ワルシャワ
    • Related Report
      2010 Final Research Report
  • [Presentation] Auto-luminescent genetically-encoded ratiometric indicator for real-time Ca2+ imaging at the single cell level2010

    • Author(s)
      Kenta Saito, Takeharu Nagai
    • Organizer
      ECS 2010 Warsaw
    • Place of Presentation
      Warsaw Ochota Campus, ワルシャワ、ポーランド
    • Related Report
      2010 Annual Research Report
  • [Presentation] Single live cell and plant Ca^<2+> imaging with BRET-based probes2008

    • Author(s)
      齊藤健太
    • Organizer
      第46回日本生物物理学会年会
    • Place of Presentation
      福岡県福岡市博多区(福岡国際会議場)
    • Year and Date
      2008-12-05
    • Related Report
      2008 Annual Research Report
  • [Remarks] ホームページ等

    • URL

      http://nano.es.hokudai.ac.jp/

    • Related Report
      2010 Final Research Report
  • [Remarks]

    • URL

      http://nano.es.hokudai.ac.jp/

    • Related Report
      2010 Annual Research Report
  • [Remarks]

    • URL

      http://nano.es.hokudai.ac.jp/paper.html

    • Related Report
      2009 Annual Research Report

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Published: 2008-04-01   Modified: 2016-04-21  

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