Development of novel mosquito-control system using Bacillus thuringiensis Cry toxins

• Project/Area Number: 24380034
• Research Category: Grant-in-Aid for Scientific Research (B)
• Allocation Type: Partial Multi-year Fund
• Section: 一般
• Research Field: Applied entomology
• Research Institution: Okayama University
• Principal Investigator: HAYAKAWA Tohru 岡山大学, 自然科学研究科, 助教 (30313555)
• Project Period (FY): 2012-04-01 – 2015-03-31
• Project Status: Completed (Fiscal Year 2014)
• Budget Amount: ¥17,420,000 (Direct Cost: ¥13,400,000、Indirect Cost: ¥4,020,000); Fiscal Year 2014: ¥4,810,000 (Direct Cost: ¥3,700,000、Indirect Cost: ¥1,110,000); Fiscal Year 2013: ¥4,810,000 (Direct Cost: ¥3,700,000、Indirect Cost: ¥1,110,000); Fiscal Year 2012: ¥7,800,000 (Direct Cost: ¥6,000,000、Indirect Cost: ¥1,800,000)
• Keywords: Bacillus thuringiensis / Mosquitocidal toxins / Cry toxin / Scorpion toxin / Synergism / Bacillus tnuringiensis / Mosquito larvae / Cry4Aa toxin / Cry11Ba toxin / Cry11Aa toxin / Cryトキシン / アカイエカ / 人工遺伝子 / 害虫防除 / 微生物殺虫剤 / Cry4Aa / Cry4Ba / Cry11Aa / Cyt1Aa / アラニンスキャニング

Outline of Final Research Achievements

Mosquitos are vectors of important diseases like malaria and viral hemorrhagic fevers. Therefore, I tried to develop novel mosquito-control system using mosquitocidal toxin derived from soil bacterium, Bacillus thuringiensis. This system were supposed to be safe and are able to prevent, or at least delay the development of resistance-insects. In this study, I established the production systems for several mosquitocidal toxins (e.g. Cry11Ba, Cyt1Aa, AahIT, BjαIT) and analyzed their mosquitocidal mechanisms by several procedures (e.g. alanine scanning, binding assay). We also observed the synergism of mosquitocidal activity between Cry4Aa and Cry11Ba.

Research Products

• [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
  • Peer Reviewed
• [Journal Article] Parasporin 1Ac2, a novel cytotoxic crystal protein isolated from Bacillus (2013)
  • Author(s): Kuroda S, Begum A, Saga M, Hirao A, Mizuki E, Sakai H, Hayakawa T.
  • Journal Title: Current microbiology Volume: 66 Issue: 5 Pages: 475-480
  • DOI: 10.1007/s00284-013-0301-1
  • Peer Reviewed
• [Journal Article] Efficient production of recombinant cystatin C using a peptide-tag, 4AaCter, that facilitates formation of insoluble protein inclusion bodies in Escherichia coli. (2013)
  • Author(s): Hayashi M, Iwamoto S, Sato S, Sudo S, Takagi M, Sakai H, Hayakawa T.
  • Journal Title: Protein Expression and Purification Volume: 88 Issue: 2 Pages: 230-234
  • DOI: 10.1016/j.pep.2013.01.011
  • Peer Reviewed
• [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
  • 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
• [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
• [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
• [Presentation] Bt菌が産生する殺蚊トキシンCry4Aaの複雑な作用機構について (2014)
  • Author(s): 早川徹、Howlader M.T.H.、中尾早織、井出徹
  • Organizer: 平成２６年度蚕糸・昆虫機能利用学術講演会
  • Place of Presentation: 神奈川県藤沢
  • Invited
• [Presentation] 殺蚊トキシンCry4Aaとサソリトキシン(Bj a IT及びAahIT)を利用する新規害虫防除法の可能性 (2013)
  • Author(s): 早川徹、松本陸、清水佳孝、酒井裕
  • Organizer: 日本蚕糸学会第83回大会
  • Place of Presentation: 筑波
  • Year and Date: 2013-03-18
• [Presentation] Functional structures of Cry4Aa toxin that are responsible for the mosquitocidal activity against Culex pipiens. (2013)
  • Author(s): Howlader M.T.H., Nakao S, Sakai H., Hayakawa T.
  • Organizer: CARES2013 International Conference on Biotechnology
  • Place of Presentation: Dhaka, Bangladesh
  • Invited
• [Presentation] Novel protein production system using a peptide-tag derived from Bacillus thuringiensis mosquitocidal Cry4Aa toxin. (2013)
  • Author(s): Hayakawa T., Sato S., Iwamoto S., Sudo S., Howlader M.T.H., Sakai H.
  • Organizer: 46th Annual Meeting of the Society for Invertebrate Pathology
  • Place of Presentation: Pittsburgh, U.S.A.
  • Invited
• [Presentation] Interactions between mosquitocidal Cry4Aa and the brush border membrane proteins of Culex pipiens larvae. (2013)
  • Author(s): Howlader M.T.H., Nakao S., Sakai H., Hayakawa T.
  • Organizer: 46th Annual Meeting of the Society for Invertebrate Pathology
  • Place of Presentation: Pittsburgh, U.S.A.
  • Invited
• [Presentation] 大腸菌を用いて殺蚊トキシンCry11Aaを効率的に生産する方法 (2013)
  • Author(s): 早川徹、佐野乙香、井出徹
  • Organizer: 第36回分子生物学会年会
  • Place of Presentation: 兵庫県神戸
  • Invited
• [Presentation] Detection of domainll loops that are responsible for mosquitocidal activity of Cry4Aa from Bacillus thuringiesis (2012)
  • Author(s): Howlader M. T. H., Nakao S., Sakai H., Hayakawa T.
  • Organizer: 第35回日本分子生物学会年会
  • Place of Presentation: 福岡
  • Year and Date: 2012-12-12
• [Presentation] 不溶性凝集体形成を促す新規ペプチドタグ(4AaCter)を利用したシスタチンCの効率的生産と精製 (2012)
  • Author(s): 林雅浩、高木麻里、佐藤真也、岩本繁久、須藤薫雄、酒井裕、早川徹
  • Organizer: 第35回日本分子生物学会年会
  • Place of Presentation: 福岡
  • Year and Date: 2012-12-12
• [Presentation] 殺蚊トキシンCryllAの受容体結合部位と考えられるドメインIIループ構造への変異導入 (2012)
  • Author(s): 金治正泰、駒田卓也、モハッマド TH ハウラダル、酒井裕、早川徹
  • Organizer: 第35回日本分子生物学会年会
  • Place of Presentation: 福岡
  • Year and Date: 2012-12-12
• [Presentation] Bt菌B0462株に由来する新規パラスポリン(PSlAc2)の解析 (2012)
  • Author(s): 早川徹、黒田将太、Begum A、佐賀瑞恵、平尾彬奈、水城英一、酒井裕
  • Organizer: 第5回パラスポリン研究会
  • Place of Presentation: 熊本
  • Year and Date: 2012-12-08
• [Book] 最新昆虫病理学、第10章 昆虫病理学の展開、４Bacillus thuringiensisの開発と利用 (2014)
  • Author(s): 早川徹、酒井裕
  • Total Pages: 251
  • Publisher: 講談社