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2003 Fiscal Year Final Research Report Summary

Kinetic analysis of phage infection towered phage therapy

Research Project

Project/Area Number 13450340
Research Category

Grant-in-Aid for Scientific Research (B)

Allocation TypeSingle-year Grants
Section一般
Research Field 生物・生体工学
Research InstitutionTokyo Institute of Technology

Principal Investigator

TANJI Yasunori  Tokyo Institute of Technology, Graduate School of Bioscience and Biotechnology, Associate Prof., 大学院・生命理工学研究科, 助教授 (00282848)

Co-Investigator(Kenkyū-buntansha) MIYANAGA Kazuhiko  Tokyo Institute of Technology, Graduate School of Bioscience and Biotechnology, Assistant Prof., 大学院・生命理工学研究科, 助手 (40323810)
HORI Kastutoshi  Tokyo Institute of Technology, Graduate School of Bioscience and Biotechnology, Assistant Prof., 大学院・生命理工学研究科, 助手 (50302956)
UNNO Hajime  Tokyo Institute of Technology, Graduate School of Bioscience and Biotechnology, Professor, 大学院・生命理工学研究科, 教授 (10087471)
Project Period (FY) 2001 – 2003
KeywordsBacteriophage / E.colt O157: H7 / Receptor / Phage-therapy / GFP / Pathogenic E.coli / Host cell recognition / Fluorcent detection
Research Abstract

T-even type coliphage PP01, which specifically infects to Escherichia coil O157: H7, uses the outer membrane protein OmpC as a receptor. The PPO01-resistant cells had lost ompC expression due to the deletion of a 14 kbp region upstream of ompC. Two host range mutants, infective to the OmpG null mutant, were isolated and gene 38, which codes for the receptor recognition protein Gp38, was sequenced from both host range mutant. According to the deduced amino acid sequence, the mutational alterations were found in Gp38. The most common mutations changed glutamine at position 161 and glycine at Dosition 208 into basic amino acids.
PP01 was also used to detect its host cell, E.coil 0157: H7. Phage capsid protein, SOC, was used as a platform to present marker protein, green fluorescent protein (GFP), on the phage capsid. DNA fragment around soc was amplified by PCR and sequenced. The gene alignment of soc and its upstream region was g56-soc.2-soc.1-soc which was the same as that of T2 phage. GFP was introduced into the C-and N-terminal regions. of SOC to produce, recombinant phages, PP01-GFP/SOC and PP01-SOC/GFP, respectively. Fusion of GFP to the SOC did not change host range of PP01. On the other hand, binding affinity of the recombinant phages to the host cell increased. However, stability of the recombinant phages in alkaline solution was reduced. Adsorption of the GFP labeled phages to the E.coli cell surface visualized the cells through fluorescent microscopy., GFP labeled PP01 adsorbed not only on the culturable E.coil cell but also viable but nonculturable (VBNC) and pasteurized cells. Coexistence of insensitive cell, E.coil K12(W3110), did not influence specificity and affinity of GFP labeled PPO01 adsorption on E coli O157: H7. GFP labeled PP01 phage could be rapid and sensitive toll of E.coil O157: H7 detection.

  • Research Products

    (19 results)

All Other

All Publications (19 results)

  • [Publications] Y.Tanji, K.Mizoguchi, M.Yoichi, M.Morita, N.Kijima, H.Kato, H.Unno: "Seasonal change and fate of coliphages infected to Escherichia coli O157:H7 in a wastewater treatment plant"Water Research. 37. 1136-1142 (2003)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] K.Mizoguchi, M.Morita, C.R.Fischer, M.Yoichi, Y Tanji, H.Unno: "The coevolution of PP01 phage and Escherichia coli O157:H7 in continuous culture"Apple.Env.Microbiol.. 69. 170-176 (2003)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] M.Oda, M.Morita, H.Unno, Y.Tanji: "Rapid Detection of Escherichia coli O157:H7 using GFP-labeled PP01 Bacteriophage"Apple.Env.Microbiol.. 70. 527-534 (2003)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Y.Tanji, T.Shimada, M.Yoichi, K.Miyanaga, K.Hori, H.Unno: "Toward rational control of Escherichia coli O157:H7 by a phage cocktail"Appl.Microbiol.Biotechnol.. (in press).

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Y.Orito, M.Morita, K.Hori, H.Unno, Y.Tanji: "Bacillus amyloliquefaciens phage endolysin can enhance permeability of Pseudomonas aeruginosa outer membrane and induce cell lysis"Appl.Microbiol.Biotechnol.. (in press).

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Y.Tanji, K.Mizoguchi, M.Yanagida, K.Hori, H.Unno: "Seasonal change of coliphage in septic tank sludge and quantification of P1 phage mediated gene transfer."J. Chem. Eng. Japan. 34. 1079-1083 (2001)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] M.Morita, K.Asami, Y.Tanji, H.Unno: "Programmed Eseherichia coil cell lysis by expression of cloned T4 phage lysis genes"Biotechnol. Prog.. 17. 573-576 (2001)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] M.Morita, Y.Tanji, K.Mizoguchi, A.Soejima, Y.Onto, H.Unno: "Antibacterial activity of Bacillus amyloliquefaciences phage endolysin without holin conjugation"J. Biosci. Bioeng.. 91. 469-474 (2001)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] M.Morita, Y Tanji, Y.Onto, K.Mizoguchi, A.Soejima, H.Unno: "Functional analysis of antibacterial activity of Bacillus amyloliquefaciens phage endolysin against Gram-negative bacteria"FEBS Letter. 500. 56-59 (2001)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] M.Morita, Y.Tanji, K.Mizoguchi, T.Akitsu, N.Kijima, H.Unno: "Characterization of virulent bacteriophage specific for Escherichia coil O157: H7 and analysis of it's cellular receptor and two tail fiber genes"FEMS Microbiol. Letter. 211. 77-83 (2002)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] M.Morita, C.R.Fischer, K.Mizoguchi, M.Yoichi, M.Oda, Y Tanji, H.Unno: "Amino acid alterations on Gp38 of host range mutant of PP11 and vidence for OmpC null mutation in its host cell, Escherichia coil O157: H7"FEMS Microbiol. Letter. 216. 243-248 (2002)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Y.Tanji, K.Mizoguchi, T.Akitsu, M.Morita, K.Hori, H.Unno.: "Fate of coliphage in waste water treatment process and detection of phages caring the shiga toxin type 2"Water Science and Technol.. 46. 285-289 (2002)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Y.Tanji, K.Mizoguchi, M.Yoichi, M.Morita, K.Hori, H.Unno: "Fate of coliphage in a wastewater treatment process"J. Biosci. Bioeng. 94. 172-174 (2002)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] C.Fisher, K.Mizoguchi, M.Morita, Y.Tanji, H.Unno.: "E.coli O157: H7can limit its susceptibility to lytic bacteriophage by excreting diffusivity-lowering factors"J. Chem. Ind. Eng.. 53. 114-115 (2002)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Y.Tanji, K.Mizoguchi, M.Yoichi, M.Morita, N.Kijima, H.Kato, H.Unno: "Seasonal change and fate of coliphages infected to Escherichia coil O157: H7 in a wastewater treatment plant"Water Research. 37. 1136-1142 (2003)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] K.Mizoguchi, M.Morita, C.R.Fischer, M.Yoichi, Y.Tanji, H.Unno: "The coevolution of PP01 phage and Escherichia coil O157: H7 in continuous culture"Apple. Env. Microbiol. 69. 170-176 (2003)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Y.Tanji, T.Shimada, M.Yoichi, K.Miyanaga, K.Hori, H.Unno.: "Toward rational control of Escherichia coil 01 57:H7 by a phage cocktail"Appl. Microbiol. Biotechnol.. (in press). DOI 10.1007/s00253-003-1438-9

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] M.Oda, M.Morita, H.Unno, Y.Tanji.: "Rapid Detection of Escherichia coil O157: H7 using GFP-labeled PP01 Bacteriophage"Apple. Env. Microbiol. 70. 527-534 (2004)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Y.Onto, M.Morita, K.Hori, H.Unno, Y.Tanji.: "Bacillus amyloliquefaciens phage endolysin can enhance permeability of Pseudomonas aeruginosa outer membrane and induce cell lysis"Appl. Microbiol. Biotechnol.. (in press). (2004)

    • Description
      「研究成果報告書概要(欧文)」より

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Published: 2005-04-19  

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