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Control of pathogen by bacteriophage carrying variable host range constructed by molecular evolution engineering

Research Project

Project/Area Number 24246133
Research Category

Grant-in-Aid for Scientific Research (A)

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

Principal Investigator

Tanji Yasunori  東京工業大学, 生命理工学院, 教授 (00282848)

Co-Investigator(Kenkyū-buntansha) 宮永 一彦  東京工業大学, 生命理工学研究科, 助教 (40323810)
Research Collaborator Hidetomo Iwano  酪農学園大学, 教授
Project Period (FY) 2012-04-01 – 2017-03-31
Project Status Completed (Fiscal Year 2016)
Budget Amount *help
¥41,600,000 (Direct Cost: ¥32,000,000、Indirect Cost: ¥9,600,000)
Fiscal Year 2016: ¥7,800,000 (Direct Cost: ¥6,000,000、Indirect Cost: ¥1,800,000)
Fiscal Year 2015: ¥7,800,000 (Direct Cost: ¥6,000,000、Indirect Cost: ¥1,800,000)
Fiscal Year 2014: ¥7,410,000 (Direct Cost: ¥5,700,000、Indirect Cost: ¥1,710,000)
Fiscal Year 2013: ¥6,760,000 (Direct Cost: ¥5,200,000、Indirect Cost: ¥1,560,000)
Fiscal Year 2012: ¥11,830,000 (Direct Cost: ¥9,100,000、Indirect Cost: ¥2,730,000)
Keywordsバクテリオファージ / ファージセラピー / 黄色ブドウ球菌 / 薬剤耐性菌 / 乳房炎 / 大腸菌 / 大腸菌O157:H7
Outline of Final Research Achievements

The emergence of antibiotic-resistant bacteria has increased the attention to the therapeutic use of bacteriophages. In order to improve the usability of phages, it is required to understand the mechanism underlying host recognition, especially receptor-binding proteins (RBPs) which determine the host range. In this study, we demonstrate staphylococcal phages ΦSA012 possesses at least two RBPs. A polyclonal antibody against ORF103 inhibited infection of ΦSA012 in the presence of α-GlcNAc, suggesting that ORF103 (ligand) binds to α-GlcNAc (Receptor).
Phage-resistant E. coli isolated from co-culture with phage T2 was used. As the target of modification, Hyper Variable Region (HVR) was selected. This region is part of gene38, which encodes the tip of the long tail fiber. CRISPR/Cas system facilitated homologous recombination between wild type phage and plasmid which encoded random mutation in the HVR. Artificially modified phage showed infectivity to the phage resistant E.coli.

Report

(6 results)
  • 2016 Annual Research Report   Final Research Report ( PDF )
  • 2015 Annual Research Report
  • 2014 Annual Research Report
  • 2013 Annual Research Report
  • 2012 Annual Research Report
  • Research Products

    (42 results)

All 2017 2016 2015 2014 2013 2012 Other

All Int'l Joint Research (2 results) Journal Article (9 results) (of which Peer Reviewed: 7 results,  Open Access: 4 results,  Acknowledgement Compliant: 2 results) Presentation (28 results) (of which Int'l Joint Research: 2 results,  Invited: 2 results) Remarks (3 results)

  • [Int'l Joint Research] The Arctic University of Norway(ノルウェー)

    • Related Report
      2016 Annual Research Report
  • [Int'l Joint Research] Ho Chi Minh City University(ベトナム)

    • Related Report
      2016 Annual Research Report
  • [Journal Article] Control of pathogen by bacteriophage2017

    • Author(s)
      Yasunori Tanji
    • Journal Title

      Biocontrol Science

      Volume: 42 Pages: 191-197

    • NAID

      40021185804

    • Related Report
      2016 Annual Research Report
  • [Journal Article] Prospective and problem of phage therapy2017

    • Author(s)
      Yasunori Tanji
    • Journal Title

      ANTIBIOTICS & CHEMOTHERAPY

      Volume: 印刷中

    • Related Report
      2016 Annual Research Report
  • [Journal Article] The presence of two receptor-binding proteins contributes to the wide host range of staphylococcal Twort-like phages2016

    • Author(s)
      Takeuchi, K. Osada, A.H. Azam, H. Asakawa, K. Miyanaga, Y. Tanji
    • Journal Title

      Appl. Environ. Microbiol.

      Volume: 82 Issue: 19 Pages: 5763-5774

    • DOI

      10.1128/aem.01385-16

    • Related Report
      2016 Annual Research Report
    • Peer Reviewed / Open Access / Acknowledgement Compliant
  • [Journal Article] Phage therapy is effective in a mouse model of bacterial equine keratitis2016

    • Author(s)
      T. Furusawa1, H. Iwano1, Y. Hiyashimizu1, K. Matsubara1, H. Higuchi, H. Nagahata, H. Niwa, Y. Katayama, Y. Kinoshita, K. Hagiwara, T. Iwasaki, Y. Tanji, H. Yokota1, Y. Tamura.
    • Journal Title

      Appl. Environ. Microbiol.

      Volume: 82 Issue: 17 Pages: 5332-5339

    • DOI

      10.1128/aem.01166-16

    • Related Report
      2016 Annual Research Report
    • Peer Reviewed / Open Access
  • [Journal Article] The Effect of Grazing on Fecal Shedding of Pathogenic Escherichia coli in Beef Cattle2015

    • Author(s)
      Sasaki H, Awais R, Takahashi J, Tanji Y, Tada C, Ogura S, Sato S, Nakai Y.
    • Journal Title

      Journal of Integrated of Integrated Field Science

      Volume: 12 Pages: 39-42

    • NAID

      120005624843

    • Related Report
      2015 Annual Research Report
    • Peer Reviewed / Open Access
  • [Journal Article] IgG-dependent aggregation of Staphylococcus aureus inhibitsbacteriophage attack2015

    • Author(s)
      Yasunori Tanji, Airi Tanaka, Kaori Tani, Miki Kurimoto, Kazuhiko Miyanaga
    • Journal Title

      Biochemical Engineering Journal

      Volume: 97 Pages: 17-24

    • DOI

      10.1016/j.bej.2015.01.007

    • Related Report
      2014 Annual Research Report
    • Peer Reviewed / Open Access / Acknowledgement Compliant
  • [Journal Article] Comparative analysis of bacterial community and antibioticresistant strains in different developmental stages of the housefly (Musca domestica)2013

    • Author(s)
      Ting Wei
    • Journal Title

      Applied Microbiology and Biotechnology

      Volume: 97 Issue: 4 Pages: 1775-1783

    • DOI

      10.1007/s00253-012-4024-1

    • Related Report
      2012 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Diffusion of bacteriophages through artificial biofilm models2012

    • Author(s)
      Jun Hu
    • Journal Title

      Biotechnology Progress

      Volume: 28 Issue: 2 Pages: 319-326

    • DOI

      10.1002/btpr.742

    • Related Report
      2012 Annual Research Report
    • Peer Reviewed
  • [Journal Article] A method for evaluating the host range of bacteriophages using phages fluorescently labeled with 5-ethynyl-2′-deoxyuridine (EdU)2012

    • Author(s)
      Sayaka Ohno
    • Journal Title

      Applied Microbiology and Biotechnology

      Volume: 95 Issue: 3 Pages: 777-788

    • DOI

      10.1007/s00253-012-4174-1

    • Related Report
      2012 Annual Research Report
    • Peer Reviewed
  • [Presentation] Application of bacteriophage and its function for controlling pathogens and expression of foreign genes on its capsid protein2016

    • Author(s)
      Yasunori Tanji, Kazuhiko Miyanaga
    • Organizer
      23rd Regional Symposium on Chemical Engineering
    • Place of Presentation
      Vung Tau City (Vietnam)
    • Year and Date
      2016-10-27
    • Related Report
      2016 Annual Research Report
    • Int'l Joint Research / Invited
  • [Presentation] ファージセラピーの可能性と問題点(牛乳房炎を例に)2016

    • Author(s)
      丹治保典
    • Organizer
      ファージ・環境ウイルス研究会 合同シンポジウム
    • Place of Presentation
      JAMSTEC横須賀本部
    • Year and Date
      2016-10-21
    • Related Report
      2016 Annual Research Report
  • [Presentation] T Genetic modification of bacteriophage by CRISPR/Cas system2016

    • Author(s)
      嶋本順明、宮永一彦、丹治保典
    • Organizer
      ファージ・環境ウイルス研究会 合同シンポジウム
    • Place of Presentation
      JAMSTEC横須賀本部
    • Year and Date
      2016-10-21
    • Related Report
      2016 Annual Research Report
  • [Presentation] Study on phage-resistant mechanism of S. aureus SA003 against phage phiSa0122016

    • Author(s)
      Aa Haeruman Azam, 宮永一彦、丹治保典
    • Organizer
      ファージ・環境ウイルス研究会 合同シンポジウム
    • Place of Presentation
      JAMSTEC横須賀本部
    • Year and Date
      2016-10-21
    • Related Report
      2016 Annual Research Report
  • [Presentation] IgG-dependent aggregation of Staphylococcus aureus inhibits bacteriophage attack2016

    • Author(s)
      丹治保典、田中愛里、谷夏織、栗本 実希、宮永一彦
    • Organizer
      第89回日本細菌学会
    • Place of Presentation
      大阪国際交流センター
    • Year and Date
      2016-03-23
    • Related Report
      2015 Annual Research Report
  • [Presentation] The host-recognition mechanism of Twort-like staphylococcal phage phiSA0122016

    • Author(s)
      竹内一平 、長田啓太、宮永一彦、丹治保典
    • Organizer
      第89回日本細菌学会
    • Place of Presentation
      大阪国際交流センター
    • Year and Date
      2016-03-23
    • Related Report
      2015 Annual Research Report
  • [Presentation] Genetic modification of bacteriophage by CRISPR/Cas system2016

    • Author(s)
      嶋本順明、星賀史也、宮永一彦、丹治保典
    • Organizer
      第89回日本細菌学会
    • Place of Presentation
      大阪国際交流センター
    • Year and Date
      2016-03-23
    • Related Report
      2015 Annual Research Report
  • [Presentation] Study on phage-resistant mechanism of S. aureus SA003 against phage phiSa0122016

    • Author(s)
      Aa Haeruman Azam、星賀史也、宮永一彦、丹治保典
    • Organizer
      第89回日本細菌学会
    • Place of Presentation
      大阪国際交流センター
    • Year and Date
      2016-03-23
    • Related Report
      2015 Annual Research Report
  • [Presentation] Coevolution between Staphylococcus aureus and its Lytic Bacteriophage ΦSA012 in Batch co-culture with Serial Transfer2015

    • Author(s)
      Yasunori Tanji
    • Organizer
      21st Biennial Evergreen International Phage Meeting
    • Place of Presentation
      The Evergreen State College
    • Year and Date
      2015-08-02
    • Related Report
      2015 Annual Research Report
    • Int'l Joint Research
  • [Presentation] ファージ研究の試行錯誤とその展望2015

    • Author(s)
      丹治保典
    • Organizer
      第2回ファージセラピー臨床応用検討会
    • Place of Presentation
      東京工業大学
    • Year and Date
      2015-07-31
    • Related Report
      2015 Annual Research Report
    • Invited
  • [Presentation] CRISPR/Casシステムによるファージゲノム編集2015

    • Author(s)
      星賀史也
    • Organizer
      第2回ファージセラピー臨床応用検討会
    • Place of Presentation
      東京工業大学
    • Year and Date
      2015-07-31
    • Related Report
      2015 Annual Research Report
  • [Presentation] 黄色ブドウ球菌特異的ファージΦSA012の宿主認識機構2015

    • Author(s)
      鈴木沙依,星賀史也,竹内一平,宮永一彦,丹治保典
    • Organizer
      第17回化学工学会学生発表会
    • Place of Presentation
      八戸高専
    • Year and Date
      2015-03-07
    • Related Report
      2014 Annual Research Report
  • [Presentation] Study on the adsorption inhibition of phage-registant Staphylococcus aureus strain SA003 against ΦSA012 phage2014

    • Author(s)
      A.H.Azam, K.Osada, K.Miyanaga, Y.Tanji
    • Organizer
      第5回ファージ研究会
    • Place of Presentation
      三重大学
    • Year and Date
      2014-09-04 – 2014-09-05
    • Related Report
      2014 Annual Research Report
  • [Presentation] 大腸菌ファージT2の宿主認識機構の解析2014

    • Author(s)
      吉崎恭平,星賀史也,宮永一彦,丹治保典
    • Organizer
      第5回ファージ研究会
    • Place of Presentation
      三重大学
    • Year and Date
      2014-09-04 – 2014-09-05
    • Related Report
      2014 Annual Research Report
  • [Presentation] 黄色ブドウ球菌のIgG依存的凝集とファージ耐性化2014

    • Author(s)
      町田桂,田中愛里,長田敬太,宮永一彦,丹治保典
    • Organizer
      第5回ファージ研究会
    • Place of Presentation
      三重大学
    • Year and Date
      2014-09-04 – 2014-09-05
    • Related Report
      2014 Annual Research Report
  • [Presentation] CRISPR/Casシステムを用いたT2ファージのゲノム改変法の開発2014

    • Author(s)
      星賀史也,髙尾信方,嶋本順明,鈴木沙依,宮永一彦,丹治保典
    • Organizer
      第5回ファージ研究会
    • Place of Presentation
      三重大学
    • Year and Date
      2014-09-04 – 2014-09-05
    • Related Report
      2014 Annual Research Report
  • [Presentation] T偶数系ファージゲノムの比較および解析2014

    • Author(s)
      宮永一彦,髙尾信方,丹治保典
    • Organizer
      第5回ファージ研究会
    • Place of Presentation
      三重大学
    • Year and Date
      2014-09-04 – 2014-09-05
    • Related Report
      2014 Annual Research Report
  • [Presentation] Aggregation of Staphylococcus aureus Which Causes Bovine Mastitis in Crude Milk2013

    • Author(s)
      Y.Tanji, K.Tani, M.Kurimoto, K.Miyanaga
    • Organizer
      The 20th Biennial International Phage Biology Meeting
    • Place of Presentation
      シアトル(USA)
    • Related Report
      2013 Annual Research Report
  • [Presentation] Comparison and Gene Analysis of T-even Phage Genomes2013

    • Author(s)
      K.Miyanaga, N.Takao, Y.Tanji
    • Organizer
      The 20th Biennial International Phage Biology Meeting
    • Place of Presentation
      シアトル(USA)
    • Related Report
      2013 Annual Research Report
  • [Presentation] T偶数系バクテリオファージにおける尾繊維タンパク質をコードする遺伝子の解析2013

    • Author(s)
      宮永一彦,丹治保典
    • Organizer
      化学工学会第45回秋季大会
    • Place of Presentation
      岡山大学
    • Related Report
      2013 Annual Research Report
  • [Presentation] 黄色ブドウ球菌とその特異的ファージΦSA012の共進化2013

    • Author(s)
      竹内一平
    • Organizer
      第15回化学工学会学生発表会
    • Place of Presentation
      山形大学(米沢市)
    • Related Report
      2012 Annual Research Report
  • [Presentation] ファージと宿主の共培養における宿主のファージ耐性化2013

    • Author(s)
      星賀史也
    • Organizer
      第15回化学工学会学生発表会
    • Place of Presentation
      山形大学(米沢市)
    • Related Report
      2012 Annual Research Report
  • [Presentation] ファージ耐性化黄色ブドウ球菌に対する再感染性ファージの解析2013

    • Author(s)
      長田 啓太
    • Organizer
      化学工学会第78年会
    • Place of Presentation
      大阪大学豊中キャンパス(豊中市)
    • Related Report
      2012 Annual Research Report
  • [Presentation] Diffusion of bacteriophages through artificial biofilm models2012

    • Author(s)
      Yasunori Tanji
    • Organizer
      Virus of microbes
    • Place of Presentation
      Brussels Belgium
    • Related Report
      2012 Annual Research Report
  • [Presentation] CONTROL OF PHASOGENS IN THE BIOFILM BY BACTERIOPHAGE2012

    • Author(s)
      Yasunori TANJI
    • Organizer
      2012 International Symposium on Advanced Biological Engineering
    • Place of Presentation
      Guilin, Guangxi, China
    • Related Report
      2012 Annual Research Report
  • [Presentation] T2ファージにおけるTail fiberの相同組換えと宿主吸着に与える影響の解析2012

    • Author(s)
      髙尾信方
    • Organizer
      第4回 ファージ研究会
    • Place of Presentation
      第4回 ファージ研究会(高崎市)
    • Related Report
      2012 Annual Research Report
  • [Presentation] IgGによる黄色ブドウ球菌凝集のファージ耐性化2012

    • Author(s)
      田中愛里
    • Organizer
      第4回 ファージ研究会
    • Place of Presentation
      第4回 ファージ研究会(高崎市)
    • Related Report
      2012 Annual Research Report
  • [Presentation] ファージセラピーに向けたファージ大量精製条件の最適化2012

    • Author(s)
      竹内一平
    • Organizer
      第4回 ファージ研究会
    • Place of Presentation
      第4回 ファージ研究会(高崎市)
    • Related Report
      2012 Annual Research Report
  • [Remarks] 東京工業大学生命理工学院 丹治研究室

    • URL

      http://www.biochemeng.bio.titech.ac.jp/index.html

    • Related Report
      2016 Annual Research Report
  • [Remarks] 東京工業大学、生命理工学院、丹治研究室ホームページ

    • URL

      http://www.biochemeng.bio.titech.ac.jp/index.html

    • Related Report
      2015 Annual Research Report
  • [Remarks] 東京工業大学大学院生命理工学研究科 丹治研究室ホームページ

    • URL

      http://www.biochemeng.bio.titech.ac.jp/index.html

    • Related Report
      2013 Annual Research Report

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Published: 2013-05-15   Modified: 2019-07-29  

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