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Novel system to produce highly functional proteins as protein inclusion in Escherichia coli.

Research Project

Project/Area Number 26660268
Research Category

Grant-in-Aid for Challenging Exploratory Research

Allocation TypeMulti-year Fund
Research Field Insect science
Research InstitutionOkayama University

Principal Investigator

Hayakawa Toru  岡山大学, 自然科学研究科, 助教 (30313555)

Project Period (FY) 2014-04-01 – 2017-03-31
Project Status Completed (Fiscal Year 2016)
Budget Amount *help
¥3,640,000 (Direct Cost: ¥2,800,000、Indirect Cost: ¥840,000)
Fiscal Year 2016: ¥1,300,000 (Direct Cost: ¥1,000,000、Indirect Cost: ¥300,000)
Fiscal Year 2015: ¥1,300,000 (Direct Cost: ¥1,000,000、Indirect Cost: ¥300,000)
Fiscal Year 2014: ¥1,040,000 (Direct Cost: ¥800,000、Indirect Cost: ¥240,000)
KeywordsBacillus thuringiensis / 4AaCter-tag / Cry4Aa / Cry46Ab / 大腸菌 / タンパク質凝集体形成 / タンパク質生産 / ペプチドタグ / 凝集体形成
Outline of Final Research Achievements

4AaCter is a peptide-tag derived from mosquitocidal Cry4Aa toxin, and fusion with this tag facilitate formation of the protein inclusion in Escherichia coli. In this study, through the analyses using various truncated mutants of 4AaCter-tag, two domains (MIF1 and 2) that related to formation of protein inclusion were identified in the amino acid stretch of 4AaCter. Our data suggested that MIF domains polymerize by binding itself and/or each other, and facilitate formation of protein inclusion. In addition, proteins (designated as P49 and P50) which may be related to the formation of protein inclusion were detected in Escherichia coli proteins. In this study, the potency of 4AaCter-tag was investigated to prepare the formulation of insecticidal Cry46Ab toxin. Application of 4AaCter-tag yielded excellent results, including increase in the production level, simplification of the product purification and high toxicity against Culex pipiens larvae.

Report

(4 results)
  • 2016 Annual Research Report   Final Research Report ( PDF )
  • 2015 Research-status Report
  • 2014 Research-status Report
  • Research Products

    (14 results)

All 2017 2016 2015 2014

All Journal Article (3 results) (of which Int'l Joint Research: 1 results,  Peer Reviewed: 2 results,  Acknowledgement Compliant: 1 results) Presentation (11 results) (of which Invited: 2 results)

  • [Journal Article] Bacillus thuringiensis Cry11Ba works synergistically with Cry4Aa but not with Cry11Aa for toxicity against mosquito Culex pipiens (Diptera: Culicidae) larvae.2017

    • Author(s)
      Hayakawa T, Yoneda N, Okada K, Higaki A, Howlader MTH, Ide T
    • Journal Title

      Applied Entomology and Zoology

      Volume: 52 Issue: 1 Pages: 61-68

    • DOI

      10.1007/s13355-016-0454-z

    • NAID

      120006406814

    • Related Report
      2016 Annual Research Report
    • Peer Reviewed / Int'l Joint Research / Acknowledgement Compliant
  • [Journal Article] 蚊を殺すトキシン由来のポリペプチドを利用して効率的なタンパク質の生産系を構築する。2016

    • Author(s)
      早川 徹
    • Journal Title

      昆虫と自然

      Volume: 51 Pages: 41-42

    • Related Report
      2016 Annual Research Report
  • [Journal Article] Potency of insect-specific scorpion toxins on mosquito control using Bacillus thur ingiensis Cry4Aa.2014

    • Author(s)
      Matsumoto R., Shimizu Y., Howlader M. T. H., Namba M., Iwamoto A., Sakai H., Hayakawa T.
    • Journal Title

      Journal of Bioscience and Bioengineering

      Volume: 117 Issue: 6 Pages: 1-4

    • DOI

      10.1016/j.jbiosc.2013.12.004

    • Related Report
      2014 Research-status Report
    • Peer Reviewed
  • [Presentation] Bacillus thuringiensisTK-E6株由来のCry46Abの殺ボウフラ活性2017

    • Author(s)
      植田翔, 黒川亜美, 槇本純, 東慶直, 早川徹, 武部聡
    • Organizer
      第61回日本応用動物昆虫学会大会
    • Place of Presentation
      東京
    • Year and Date
      2017-03-27
    • Related Report
      2016 Annual Research Report
  • [Presentation] ペプチドタグ4AaCterを利用したCryトキシンの製剤化2017

    • Author(s)
      早川徹、岡崎友亮、井出徹
    • Organizer
      平成29年度蚕糸・昆虫機能利用学術講演会
    • Place of Presentation
      茨城県つくば市
    • Year and Date
      2017-03-21
    • Related Report
      2016 Annual Research Report
  • [Presentation] 抵抗性の発達阻止を可能とする新しい蚊防除資材2016

    • Author(s)
      早川徹、榊原暁、植田翔、武部聡
    • Organizer
      第12回昆虫病理研究会シンポジウム
    • Place of Presentation
      宮城県岩沼市
    • Year and Date
      2016-09-15
    • Related Report
      2016 Annual Research Report
    • Invited
  • [Presentation] ボウフラに対して食毒性を示す非3-ドメインCry型33-kDaタンパク質2016

    • Author(s)
      植田翔、根来綾子、大田雅俊、黒川亜美、槇本純、東慶直、早川徹、武部聡
    • Organizer
      第12回昆虫病理研究会シンポジウム
    • Place of Presentation
      宮城県岩沼市
    • Year and Date
      2016-09-15
    • Related Report
      2016 Annual Research Report
  • [Presentation] 殺蚊Cryトキシンが示す殺虫活性助長作用2016

    • Author(s)
      早川徹、迫田陽子、米田直也、井出徹
    • Organizer
      平成28年度蚕糸・昆虫機能利用学術講演会-日本蚕糸学会第86回大会-
    • Place of Presentation
      京都
    • Year and Date
      2016-03-17
    • Related Report
      2015 Research-status Report
  • [Presentation] Bt菌が産生するCryトキシンを用いて蚊の効率的な防除法を開発する試み2015

    • Author(s)
      早川徹、迫田陽子、米田直也、井出徹
    • Organizer
      第38回日本分子生物学会年会、第88回日本生化学会大会 合同大会
    • Place of Presentation
      神戸
    • Year and Date
      2015-12-01
    • Related Report
      2015 Research-status Report
  • [Presentation] 殺蚊トキシン由来のペプチドタグ4AaCterによるタンパク質凝集体形成機構2015

    • Author(s)
      早川徹、岡崎友亮、中西里菜、永尾権、佐藤友美、井出徹
    • Organizer
      平成27年度 蚕糸・昆虫機能利用学術講演会-日本蚕糸学会第85回大会-
    • Place of Presentation
      札幌
    • Year and Date
      2015-09-26
    • Related Report
      2015 Research-status Report
  • [Presentation] 効率的且つ持続的蚊防除システムの構築に向けた最近の取り組み2014

    • Author(s)
      早川徹
    • Organizer
      第11回昆虫病理研究会シンポジウム
    • Place of Presentation
      山梨県富士吉田
    • Year and Date
      2014-09-18 – 2014-09-20
    • Related Report
      2014 Research-status Report
    • Invited
  • [Presentation] 殺蚊トキシンCry4Aaの作用機構に関する研究2014

    • Author(s)
      岡田光司、Howlader MTH、中尾早織、井出徹、早川徹
    • Organizer
      第11回昆虫病理研究会シンポジウム
    • Place of Presentation
      山梨県富士吉田
    • Year and Date
      2014-09-18 – 2014-09-20
    • Related Report
      2014 Research-status Report
  • [Presentation] 殺蚊トキシンCry11Baを用いた効率的蚊防除システムの構築を目指して2014

    • Author(s)
      米田直也、岡田光司、檜垣亜由子、井出徹、早川徹
    • Organizer
      第11回昆虫病理研究会シンポジウム
    • Place of Presentation
      山梨県富士吉田
    • Year and Date
      2014-09-18 – 2014-09-20
    • Related Report
      2014 Research-status Report
  • [Presentation] BT菌亜種israelensisに由来するCyt1Aaの大腸菌を用いた効率的生産法2014

    • Author(s)
      檜垣亜由子、岡田光司、米田直也、井出徹、早川徹
    • Organizer
      第11回昆虫病理研究会シンポジウム
    • Place of Presentation
      山梨県富士吉田
    • Year and Date
      2014-09-18 – 2014-09-20
    • Related Report
      2014 Research-status Report

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Published: 2014-04-04   Modified: 2018-03-22  

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