Realization of a secure and safe proteinaceous-insecticide based on the directed evolution of the insecticidal toxin from soil bacterium, Bacillus thuringiensis

• Project/Area Number: 21310051
• Research Category: Grant-in-Aid for Scientific Research (B)
• Allocation Type: Single-year Grants
• Section: 一般
• Research Field: Environmental technology/Environmental materials
• Research Institution: Tokyo University of Agriculture and Technology
• Principal Investigator: SATO Ryoichi 東京農工大学, 大学院・農学研究院, 教授 (30235428)
• Co-Investigator(Kenkyū-buntansha): KUNIMI Yasuhisa 東京農工大学, 大学院・農学研究院, 教授 (50195476)
• Project Period (FY): 2009 – 2011
• Project Status: Completed (Fiscal Year 2011)
• Budget Amount: ¥18,460,000 (Direct Cost: ¥14,200,000、Indirect Cost: ¥4,260,000); Fiscal Year 2011: ¥5,070,000 (Direct Cost: ¥3,900,000、Indirect Cost: ¥1,170,000); Fiscal Year 2010: ¥5,070,000 (Direct Cost: ¥3,900,000、Indirect Cost: ¥1,170,000); Fiscal Year 2009: ¥8,320,000 (Direct Cost: ¥6,400,000、Indirect Cost: ¥1,920,000)
• Keywords: 環境負荷低減技術 / 殺虫剤 / Cry toxin / 進化分子工学 / Bacillus thuiringiensis / タンパク質殺虫剤 / カドヘリン様受容体 / Bacillus thuringiensis

Research Abstract

It was indicated that Cry toxin mutants affinity-maturated to cadherin-like receptor is expected to be easily acquired, if mutation is introduced in loop 2 or loop 3 of Cry toxin, mutant Cry toxins are displayed on the T7 phage, and mutant-toxin-displaying phages are screened by bio-panning using cadherin-like receptor. That is to say, base technology was developed for the directed evolution of Cry toxin by which any available Cry toxins are possible to be changed to be highly active to any insect pests

Research Products

• [Journal Article] Response of midgut epithelial cells to Cry1Aa is toxin-dependent and depends on the interplay between toxic action and the host apoptotic response (2012)
  • Author(s): Tanaka S, Yoshizawa Y, Sato R
  • Journal Title: FEBS Journal Volume: 279 Issue: 6 Pages: 1071-1079
  • DOI: 10.1111/j.1742-4658.2012.08499.x
  • Peer Reviewed
• [Journal Article] Bacillus thuringiensis Cry toxins bound specifically to various proteins via domain III, which had a galactose-binding domain-like fold (2011)
  • Author(s): Kitami M, Kadotani T, Nakanishi K, Atsumi S, Higurashi S, Ishizaka T, Watanabe A, Sato R
  • Journal Title: Bioscience Biotechnology Biochemistry Volume: 75 Pages: 305-312
  • NAID: 10027897803
  • URL: http://www.jstage.jst.go.jp/article/bbb/75/2/75_100689/_article
  • Peer Reviewed
• [Journal Article] Bacillus thuringiensis Cry Toxins Bound Specifically to Various Proteins via Domain III, Which Had a Galactose-Binding Domain-Like Fold. (2011)
  • Author(s): M.Kitami, et al.
  • Journal Title: Biosci Biotechnol Biochem Volume: 75(2) Pages: 305-312
  • Peer Reviewed
• [Journal Article] Analysis of the region for receptor binding and triggering of oligomerization on Bacillus thuringiensis Cry1Aa toxin (2009)
  • Author(s): Obata F, Kitami M, Inoue Y, Atsumi S, Yoshizawa Y, Sato R
  • Journal Title: FEBS Journal Volume: 276 Issue: 20 Pages: 5949-5959
  • DOI: 10.1111/j.1742-4658.2009.07275.x
  • Peer Reviewed
• [Journal Article] Analysis of the region for receptor binding and triggering of oligomer ization on Bacillus thuringiensis CrylAa toxin. (2009)
  • Author(s): F.Obata, et al.
  • Journal Title: FEBS Journal 276 Pages: 5949-5959
  • Peer Reviewed
• [Presentation] バイオパニング法を用いて選抜されたBtR175高親和性Cry1Aa変異体の解析 (2011)
  • Author(s): 藤井勇樹, 他3名
  • Organizer: 第73回昆虫病理研究会
  • Place of Presentation: 東京大学(弥生)
  • Year and Date: 2011-12-03
• [Presentation] Cry1A毒素に対する宿主昆虫感受性の決定に関与する2分子の解析 (2011)
  • Author(s): 田中詩穂, 他3名
  • Organizer: 第73回昆虫病理研究会
  • Place of Presentation: 東京大学(弥生)
  • Year and Date: 2011-12-03
• [Presentation] Bt殺虫性毒素の作用機構 (2011)
  • Author(s): 佐藤令一
  • Organizer: 第84回日本生化学会大会 シンポジウム2S3a
  • Place of Presentation: 国立京都国際会館(招待講演)
  • Year and Date: 2011-09-22
• [Presentation] Bt殺虫性毒素の作用機構 (2011)
  • Author(s): 佐藤令一
  • Organizer: 第84回日本生化学会大会(招待講演)
  • Place of Presentation: 国立京都国際会館
  • Year and Date: 2011-09-11
• [Presentation] Tow molecular components from the midgut cells of Bombyx mori required for making Sf9 cell susceptible to Cry1A toxins (2011)
  • Author(s): Tanaka S, Miyamoto K, Noda H, Sato R
  • Organizer: 第84回日本生化学会大会
  • Place of Presentation: 国立京都国際会館
  • Year and Date: 2011-09-11
• [Presentation] Analysis on the role of Bacillus thuringiensis Cry toxin loop regions in binding affinity to the cadherin-like receptor using Cry toxin mutants (2011)
  • Author(s): Fujii Y, Kotani T, Morimoto C, Harashima Y, Hoshino Y, Sato R
  • Organizer: 44TH Annual Meeting of the Society for Invertebrate Pathology
  • Place of Presentation: Halifax, Canada
  • Year and Date: 2011-08-08
• [Presentation] Variation of host cell components required for toxic actions of different subclasses of Cry1 toxin (2011)
  • Author(s): Shiho S, Tsukamoto H, Miyamoto K, Noda H, Sato R
  • Organizer: 44TH Annual Meeting of the Society for Invertebrate Pathology
  • Place of Presentation: Halifax, Canada
  • Year and Date: 2011-08-08
• [Presentation] Binding affinity improvement of the Cry1Aa toxin to the cadherin-like protein, BtR175 from Bombyx mori using phage display of loop 3 mutant toxins and bio-panning (2011)
  • Author(s): Fujii Y, Otsuki M, Hoshino Y, Sato R
  • Organizer: 44TH Annual Meeting of the Society for Invertebrate Pathology
  • Place of Presentation: Halifax, Canada
  • Year and Date: 2011-08-08
• [Presentation] Bombyx mori midgut epithelial cells' reaction induced by Cry1Aa includes both or either of osmotic swelling and apoptosis depending on toxic condition (2011)
  • Author(s): Tanaka S, Yoshizawa Y. Sato R
  • Organizer: 44TH Annual Meeting of the Society for Invertebrate Pathology
  • Place of Presentation: Halifax, Canada
  • Year and Date: 2011-08-08
• [Presentation] Binding affinity improvement of the Cry1Aa toxin to the cadherin-like protein, BtR175 from Bombyx mori using phage display of loop 3 mutant (2011)
  • Author(s): Yuki Fujii, 他3名
  • Organizer: 44TH Annual Meeting of the Society for Invertebrate Pathology
  • Place of Presentation: Halifax, Canada
  • Year and Date: 2011-08-08
• [Presentation] Bombyx mori midgut epithelial cells' reaction induced by Cry1Aa includes both or either of osmotic swelling and apoptosis depending on toxic (2011)
  • Author(s): Shiho Tanaka, 他2名
  • Organizer: 44TH Annual Meeting of the Society for Invertebrate Pathology
  • Place of Presentation: Halifax, Canada
  • Year and Date: 2011-08-08
• [Presentation] カイコガのカドヘリン様受容体に対して結合和性が向上したCry1Aa異の作製法 (2011)
  • Author(s): 大槻愛美, 他3名
  • Organizer: 平成23年度蚕糸・昆虫機能利用学術講演会
  • Place of Presentation: 日本蚕糸学会第81回大会講演要旨集
  • Year and Date: 2011-03-20
• [Presentation] 非感受性昆虫に活性を獲得した Cry毒素変異体の取得法の構築 (2011)
  • Author(s): 小山祐美, 佐藤令一
  • Organizer: 平成23年度蚕糸・昆虫機能利用学術講演会
  • Place of Presentation: 日本蚕糸学会第81回大会講演要旨集
  • Year and Date: 2011-03-20
• [Presentation] 変異体を用いたBt菌Cry毒素のループ部位の受容体結合和性における役割の解析 (2010)
  • Author(s): 藤井勇樹, 他5名
  • Organizer: 第33回日本分子生物学会年会
  • Place of Presentation: 神戸ポートアイランド
  • Year and Date: 2010-12-08