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Development of phage surface engineering for green biotechnology

Research Project

Project/Area Number 16360408
Research Category

Grant-in-Aid for Scientific Research (B)

Allocation TypeSingle-year Grants
Section一般
Research Field Biofunction/Bioprocess
Research InstitutionTokyo Institute of Technology

Principal Investigator

TANJI Yasunori  Tokyo Inst. Of Technol., Dept.Bioeng., Associate Prof., 大学院生命理工学研究科, 助教授 (00282848)

Co-Investigator(Kenkyū-buntansha) MIYANAGA Kazuhiko  Tokyo Inst. Of Technol., Dept.Bioeng., Assistant Prof., 大学院生命理工学研究科, 助手 (40323810)
海野 肇  東京工業大学, 大学院・生命理工学研究科, 教授 (10087471)
Project Period (FY) 2004 – 2006
Project Status Completed (Fiscal Year 2006)
Budget Amount *help
¥12,500,000 (Direct Cost: ¥12,500,000)
Fiscal Year 2006: ¥2,700,000 (Direct Cost: ¥2,700,000)
Fiscal Year 2005: ¥3,300,000 (Direct Cost: ¥3,300,000)
Fiscal Year 2004: ¥6,500,000 (Direct Cost: ¥6,500,000)
Keywordsgreen chemistry / bacteriophage / surface engineering / Escherichia coli / 緑色蛍光タンパク質 / キトサナーゼ
Research Abstract

We designed a bacteriophage T2 system to display proteins fused at the N terminus of the head protein small outer capsid (SOC) of T2 phage. To facilitate selection of chimeric phage, a T2 phage encoding the β-galactosidase gene (βgal) upstream of the soc gene was constructed. The phage, named T2βGal, produces blue plaques on agar plates containing XGal. Subsequently, a plasmid encoding the target protein upstream of soc was constructed and used to transform E.coli B^E cells. Transformed cells were infected with T2βGal and homologous recombination between phage DNA and the plasmid resulted in a chimeric phage which produced transparent plaques due to the excision of the βgal gene. Chitosanase of Bacillus sp. strain K17 (ChoK), consisting of 453 amino acids, was used as a model target protein. Recombinant T2 phage which produced ChoK was named T2ChoK. T2ChoK was produced from T2βGal at a recombination frequency of about 0.1 %. On the other hand, the value for T2IβGal produced from wild type T2 was 0.001 %. This new system enables us to select recombinant phage very quickly and accurately. The number of molecules of ChoK was calculated at 14.7 per single phage. Latent period, and burst size were estimated for chimeric phages.

Report

(4 results)
  • 2006 Annual Research Report   Final Research Report Summary
  • 2005 Annual Research Report
  • 2004 Annual Research Report
  • Research Products

    (18 results)

All 2006 2005 2004

All Journal Article (16 results) Book (1 results) Patent(Industrial Property Rights) (1 results)

  • [Journal Article] A recombinant bacteriophage-based assay for the discriminative detection of culturable and viable but Nonculturable Escherichi coli O157 : H7.2006

    • Author(s)
      Raheela, H., Fukudomi.K., Miyanaga, H., Unno, Y., Tanji
    • Journal Title

      Biotechnol. Prog. 22

      Pages: 853-859

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2006 Final Research Report Summary
  • [Journal Article] Detection of Escherichia coli in the sewage influent by fluorescent labeled T4 phage.2006

    • Author(s)
      K.Miyanaga, T.Hijikata, C.Furukawa, H.Unno, Y.Tanji.
    • Journal Title

      Biochem. Eng. J. 29

      Pages: 119-124

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2006 Annual Research Report 2006 Final Research Report Summary
  • [Journal Article] A recombinant bacteriophage-based assay for the discriminative detection of culturable and viable but Nonculturable Escherichi coli 0157 : H72006

    • Author(s)
      Raheela, H.Fukudomi.K.Miyanaga, H.Unno, Y.Tanji
    • Journal Title

      Biotechnol.Prog. 22

      Pages: 853-859

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2006 Final Research Report Summary
  • [Journal Article] Detection of Escherichia coli in the sewage influent by fluorescent labeled T4 phage.2006

    • Author(s)
      K.Miyanaga, T.Hijikata, C.Furukawa, H.Unno, Y.Tanji.
    • Journal Title

      Biochem.Eng.J. 29

      Pages: 119-124

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2006 Final Research Report Summary
  • [Journal Article] A recombinant bacteriophage-based assay for the discriminative detection of culturable and viable but Noncultruable Escherichi coli O157 : H72006

    • Author(s)
      Reheela, H.Fukudomi., K.Miyanaga, H.Unno, Y.Tanji
    • Journal Title

      Biotechnol. Prog. 22

      Pages: 853-859

    • Related Report
      2006 Annual Research Report
  • [Journal Article] Therapeutic use of Phage Cocktail for Controlling Escherichia coli O157:H7 in Gastrointestinal Tract of Mice.2005

    • Author(s)
      Y.Tanji, T.Shimada, H.Fukudomi, K.Miyanaga, Y.Nakai, H.Unno
    • Journal Title

      J.Biosci.Bioeng.,100,280-287,(2005) 100

      Pages: 280-287

    • NAID

      210000130371

    • Related Report
      2005 Annual Research Report
  • [Journal Article] Quick Selection of Chimeric T2 Phage Which Displays Active Enzyme on the Viral Capsid2005

    • Author(s)
      Y.Tanji, K.Murofushi, K.Miyanaga
    • Journal Title

      Biotechnol.Progress 21

      Pages: 1768-1771

    • Related Report
      2005 Annual Research Report
  • [Journal Article] バクテリオファージの科学と応用(その1)、ファージの特長と自然界における役割2005

    • Author(s)
      丹治保典
    • Journal Title

      バイオインダストリー 22(1)

      Pages: 66-67

    • Related Report
      2005 Annual Research Report
  • [Journal Article] バクテリオファージの科学と応用(その2)、ファージセラピー2005

    • Author(s)
      丹治保典
    • Journal Title

      バイオインダストリー 22(2)

      Pages: 78-80

    • Related Report
      2005 Annual Research Report
  • [Journal Article] バクテリオファージの科学と応用(その3)、ファージ利用の新展開2005

    • Author(s)
      丹治保典
    • Journal Title

      バイオインダストリー 22(3)

      Pages: 564-565

    • Related Report
      2005 Annual Research Report
  • [Journal Article] Alteration of tail fiber protein gp38 enables T2 phage to infect Escherichia coli O157 : H72005

    • Author(s)
      M.Yoichi, M.Abe, K.Miyanaga, H.Unno, Y.Tanji
    • Journal Title

      J.Biotechnol. 115

      Pages: 101-107

    • Related Report
      2004 Annual Research Report
  • [Journal Article] The coexistence of Escherichia coli serotype O157 : H7 and its specific bacteriophage in continuous culture2004

    • Author(s)
      C.R.Fischer, M.Yoichi, H.Unno, Y.Tanji
    • Journal Title

      FEMS Microbiol.Letter 241

      Pages: 171-177

    • NAID

      120006581737

    • Related Report
      2004 Annual Research Report
  • [Journal Article] Escherichia coli detection by GFP-labeled lysozyme-inactivated T4 Bacteriophage.2004

    • Author(s)
      Y.Tanji, C.Furukawa, N.Suk-Hyun, T.Hijikata, K.Miyanaga, H.
    • Journal Title

      J.Biotechnol. 114

      Pages: 11-20

    • Related Report
      2004 Annual Research Report
  • [Journal Article] Toward rational control of Escherichia coli O157 : H7 by a phage cocktail2004

    • Author(s)
      Y.Tanji, T.Shimada, M.Yoichi, K.Miyanaga, K Hori, H.Unno
    • Journal Title

      Appl.Microbiol.Biotechnol. 64(2)

      Pages: 270-274

    • Related Report
      2004 Annual Research Report
  • [Journal Article] The criterion for selecting effective phage for Escherichia coli O157 : H7 control2004

    • Author(s)
      M.Yoichi, M.Marita, K.Mizoguchi, C.R.Fischer, H.Unno, Y.Tanji
    • Journal Title

      Biochemical Engineering J. 19

      Pages: 221-227

    • Related Report
      2004 Annual Research Report
  • [Journal Article] Bacillus amyloliquefaciens phage endolysin can enhance permeability of Pseudomonas aeruginosa outer membrane and induce cell lysis2004

    • Author(s)
      Y.Orito, M.Morita, K.Hori, H.Unno, Y.Tanj
    • Journal Title

      Appl.Microbiol.Biotechnol. 65

      Pages: 105-109

    • Related Report
      2004 Annual Research Report
  • [Book] バイオプロダクション(細菌の死の定義と汚泥減容化)2006

    • Author(s)
      丹治保典(共著)
    • Publisher
      コロナ社
    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2006 Annual Research Report 2006 Final Research Report Summary
  • [Patent(Industrial Property Rights)] ファージ頭殻提示腸管免疫誘導型ワクチン2006

    • Inventor(s)
      丹治 保典, 中井 裕, 他2名
    • Industrial Property Rights Holder
      TLO
    • Industrial Property Number
      2006-024518
    • Filing Date
      2006-02-01
    • Related Report
      2005 Annual Research Report

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Published: 2004-04-01   Modified: 2016-04-21  

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