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Development of a novel protein purification/regulation methodology by using a temperature-sensitive monoclonal antibody.

Research Project

Project/Area Number 26650024
Research Category

Grant-in-Aid for Challenging Exploratory Research

Allocation TypeMulti-year Fund
Research Field Structural biochemistry
Research InstitutionWaseda University

Principal Investigator

Okano Toshiyuki  早稲田大学, 理工学術院, 教授 (40272471)

Project Period (FY) 2014-04-01 – 2016-03-31
Project Status Completed (Fiscal Year 2015)
Budget Amount *help
¥3,900,000 (Direct Cost: ¥3,000,000、Indirect Cost: ¥900,000)
Fiscal Year 2015: ¥1,950,000 (Direct Cost: ¥1,500,000、Indirect Cost: ¥450,000)
Fiscal Year 2014: ¥1,950,000 (Direct Cost: ¥1,500,000、Indirect Cost: ¥450,000)
Keywordsモノクローナル抗体 / タンパク質相互作用 / 一本鎖抗体 / サーモジェネティクス / タンパク質間相互作用
Outline of Final Research Achievements

We recently developed a monoclonal antibody that shows strong temperature dependence in the binding to the epitope peptide, a carboxyl-terminal sequence of chicken CRY4.
In this study, we tested the possibility to use this epitope peptide as a tag for temperature-dependent affinity purification of the tagged-proteins. We found the binding of the antibody with epitope-tagged glutathione S-transferase (GST) and yellow fluorescent protein (YFP) in a temperature-dependent manner. We also succeeded in the temperature-dependent purification of those proteins.
This technology is the establishment of a new tag and purification methods of proteins, and also it would be applicable in a various fields to the temperature-dependent control of protein localization and function.

Report

(3 results)
  • 2015 Annual Research Report   Final Research Report ( PDF )
  • 2014 Research-status Report
  • Research Products

    (18 results)

All 2016 2015 2014

All Journal Article (3 results) (of which Peer Reviewed: 3 results,  Open Access: 2 results,  Acknowledgement Compliant: 3 results) Presentation (14 results) (of which Int'l Joint Research: 1 results,  Invited: 3 results) Book (1 results)

  • [Journal Article] Rhodopsin in the Dark Hot Sea: Molecular Analysis of Rhodopsin in a Snailfish, Careproctus rhodomelas, Living near the Deep-Sea Hydrothermal Vent2015

    • Author(s)
      Rie Sakata, Ryo Kabutomori, Keiko Okano, Hiromasa Mitsui, Akihiro Takemura, Tetsuya Miwa, Hiroyuki Yamamoto, Toshiyuki Okano
    • Journal Title

      PLoS ONE

      Volume: 10 Issue: 8 Pages: e0135888-e0135888

    • DOI

      10.1371/journal.pone.0135888

    • Related Report
      2015 Annual Research Report
    • Peer Reviewed / Open Access / Acknowledgement Compliant
  • [Journal Article] Overexpression in yeast, photocycle, and in vitro structural change of an avian putative magnetoreceptor cryptochrome42015

    • Author(s)
      Hiromasa Mitsui, Toshinori Maeda, Chiaki Yamaguchi, Yusuke Tsuji, Ryuji Watari, Yoko Kubo, Keiko Okano, and Toshiyuki Okano
    • Journal Title

      Biochemistry

      Volume: 54 Issue: 10 Pages: 1908-1917

    • DOI

      10.1021/bi501441u

    • Related Report
      2014 Research-status Report
    • Peer Reviewed / Acknowledgement Compliant
  • [Journal Article] Hypothalamic Expression and Moonlight-Independent Changes of Cry3 and Per4 Implicate Their Roles in Lunar Clock Oscillators of the Lunar-Responsive Goldlined Spinefoot2014

    • Author(s)
      Riko Toda, Keiko Okano, Yuki Takeuchi, Chihiro Yamauchi, Masato Fukushiro, Akihiro Takemura, Toshiyuki Okano
    • Journal Title

      PLoS ONE

      Volume: 9 Issue: 10 Pages: e109119-e109119

    • DOI

      10.1371/journal.pone.0109119

    • Related Report
      2014 Research-status Report
    • Peer Reviewed / Open Access / Acknowledgement Compliant
  • [Presentation] Biological Rhythms and Cryptochromes in Xenopus tropicalis2016

    • Author(s)
      Toshiyuki OKANO
    • Organizer
      International Meeting on Aquatic Model Organisms for Human Disease and Toxicology Research
    • Place of Presentation
      Okazaki Conference Center
    • Year and Date
      2016-03-18
    • Related Report
      2015 Annual Research Report
    • Int'l Joint Research / Invited
  • [Presentation] ドジョウ(Misgurnus anguillicaudatus)の末梢組織における時計遺伝子の光同調2016

    • Author(s)
      更谷有哉、竹内悠記、岡野恵子、岡野俊行
    • Organizer
      日本動物学会関東支部
    • Place of Presentation
      神奈川大学
    • Year and Date
      2016-03-12
    • Related Report
      2015 Annual Research Report
  • [Presentation] 月周性産卵魚類ゴマアイゴの脳内時計遺伝子発現に月光が与える影響2016

    • Author(s)
      兜森椋、竹内悠記、山内千裕、宮城ひとみ、竹村明洋、岡野恵子、岡野俊行
    • Organizer
      日本動物学会関東支部
    • Place of Presentation
      神奈川大学
    • Year and Date
      2016-03-12
    • Related Report
      2015 Annual Research Report
  • [Presentation] 温度依存的に親和性を変化させるモノクローナル抗体を利用したタンパク質精製・動態制御系の構築2015

    • Author(s)
      大嶋 拓哉、三浦 宏太、熨斗 洋星、岡野 恵子、岡野 俊行
    • Organizer
      日本分子生物学会
    • Place of Presentation
      神戸
    • Year and Date
      2015-12-01
    • Related Report
      2015 Annual Research Report
  • [Presentation] ドジョウCryファミリー遺伝子の同定と発現解析2015

    • Author(s)
      竹内 悠記、更谷 有哉、阿部 大輝、岡野 恵子、岡野 俊行
    • Organizer
      日本分子生物学会
    • Place of Presentation
      神戸
    • Year and Date
      2015-12-01
    • Related Report
      2015 Annual Research Report
  • [Presentation] ゼブラフィッシュにおけるクリプトクロムの発現変動解析2015

    • Author(s)
      玉澤歩実、岡野恵子、戸田りこ、岡野俊行
    • Organizer
      日本時間生物学会
    • Place of Presentation
      東京
    • Year and Date
      2015-11-21
    • Related Report
      2015 Annual Research Report
  • [Presentation] ドジョウにおけるCry1a,cの同定と発現解析2015

    • Author(s)
      更谷有哉、竹内悠記、阿部大輝、岡野恵子、岡野俊行
    • Organizer
      日本時間生物学会
    • Place of Presentation
      東京
    • Year and Date
      2015-11-21
    • Related Report
      2015 Annual Research Report
  • [Presentation] フグ眼球由来培養細胞における時計関連遺伝子の光応答性2015

    • Author(s)
      伊藤正晴、岡野恵子、小澤翔一、佐藤駿、小太刀佐和、宮台俊明、竹村明洋、岡野俊行
    • Organizer
      日本時間生物学会
    • Place of Presentation
      東京
    • Year and Date
      2015-11-21
    • Related Report
      2015 Annual Research Report
  • [Presentation] ネッタイツメガエルの光と時間の生物学2015

    • Author(s)
      岡野俊行
    • Organizer
      日本動物学会
    • Place of Presentation
      新潟
    • Year and Date
      2015-09-17
    • Related Report
      2015 Annual Research Report
    • Invited
  • [Presentation] ニワトリクリプトクロム4の光反応の特性解析2015

    • Author(s)
      MITSUI Hiromasa, OKANO Keiko, OKANO Toshiyuki
    • Organizer
      生物物理学会
    • Place of Presentation
      金沢
    • Year and Date
      2015-09-13
    • Related Report
      2015 Annual Research Report
  • [Presentation] Pichia Pastorisによるニワトリクリプトクロム4の発現系の構築と分光学的解析2015

    • Author(s)
      酒井一輝, 三井広大, 岡野恵子, 岡野俊行
    • Organizer
      日本動物学会
    • Place of Presentation
      早稲田大学
    • Year and Date
      2015-03-14
    • Related Report
      2014 Research-status Report
  • [Presentation] CRIP4との相互作用におけるニワトリCRY1のヘム調節モチーフの役割2014

    • Author(s)
      大塚裕介、葛川純也、岡野恵子、岡野俊行
    • Organizer
      日本時間生物学会
    • Place of Presentation
      九州大学
    • Year and Date
      2014-11-08 – 2014-11-09
    • Related Report
      2014 Research-status Report
  • [Presentation] 脊椎動物クリプトクロムの多様性、機能および分子応用2014

    • Author(s)
      岡野俊行
    • Organizer
      日本生物物理学会
    • Place of Presentation
      札幌コンベンションセンター
    • Year and Date
      2014-09-26
    • Related Report
      2014 Research-status Report
    • Invited
  • [Presentation] ニワトリクリプトクロム4の光反応特性に外部環境が与える影響2014

    • Author(s)
      三井広大, 前田俊徳, 山口千秋, 辻悠佑, 酒井一輝, 岡野恵子, 岡野俊行
    • Organizer
      日本生物物理学会
    • Place of Presentation
      札幌コンベンションセンター
    • Year and Date
      2014-09-25 – 2014-09-27
    • Related Report
      2014 Research-status Report
  • [Book] 研究者が教える動物実験 第一巻 感覚(分担)2015

    • Author(s)
      岡野俊行、岡野恵子
    • Total Pages
      4
    • Publisher
      共立出版
    • Related Report
      2015 Annual Research Report

URL: 

Published: 2014-04-04   Modified: 2017-05-10  

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