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Comparative genome analysis & enterohemorrhagic Escherichia coli O157 and its clinical application.

Research Project

Project/Area Number 13470061
Research Category

Grant-in-Aid for Scientific Research (B)

Allocation TypeSingle-year Grants
Section一般
Research Field Bacteriology (including Mycology)
Research InstitutionMiyazaki Medical College

Principal Investigator

HAYASHI Tetsuya  Miyazaki Medical College, Faculty of Medicine, Professor, 医学部, 教授 (10173014)

Co-Investigator(Kenkyū-buntansha) NAKAYAMA Keisuke  Miyazaki Medical College, Faculty Of Medicine, Research Associate, 医学部, 助手 (10347057)
OHNISHI Makoto  Miyazaki Medical College, Faculty of Medicine, Associate Professor, 医学部, 助教授 (10233214)
Project Period (FY) 2001 – 2002
Project Status Completed (Fiscal Year 2002)
Budget Amount *help
¥12,500,000 (Direct Cost: ¥12,500,000)
Fiscal Year 2002: ¥3,700,000 (Direct Cost: ¥3,700,000)
Fiscal Year 2001: ¥8,800,000 (Direct Cost: ¥8,800,000)
Keywordsenterohemorrhagic Escherichia coli O157 / genome diversity / bacteriophage / PCR / strain-discrimination system
Research Abstract

The aims of the project were to reveal the genomic diversity of Escherichia coli O157, and to develop a new tool for molecular epidemiological studies of O157 infection based on the genomic diversity.
We first selected eight O157 strains that exhibited diverse Xbal-digestion patterns from 1796 strains isolated in Japan in 1998. Next, based on the genome sequence of O157 Sakai, we prepared a set of PCR primers to amplify 560 DNA segments covering the whole genome of O157 (the average length of segments was about 10 Kb). By using long PCR and the primer set, we analyzed the whole genome structures of the eight O157 strains using the Sakai strain as a reference. By this analysis which we named Whole Genome PCR Scanning (WGPS), we could identify all the genomic regions with any structural polymorphism, indicating that WGPS is a powerful technique for the comparative analysis of closely related genomes. The data also indicate that an unexpectedly high degree of genomic diversity exists among O157 strains. In particular, prophage regions, including Stx-transducing phages, exhibited extensive diversities.
By the WGPS analysis, about 30 genomic regions exhibiting small-size structural polymorphisms detectable by conventional PCR were also identified. Selecting 9 target regions among them, we constructed a multiplex PCR system. Since the stx1, stx2 and eae genes were also included as targets, the system was finally composed of 12 PCR reactions. In our preliminary examination, this system classified 51 strains into 21 types, suggesting that the system could be developed as a useful tool for the epidemiological studies, but some improvement is still required.
In addition, we performed comparative and functional analyses of some O157 genes, and found that urease genes are specifically distributed to strains belonging to enterohemorrhagic E. coli, and that the toxB gene is involved in epithelial cell adherence.

Report

(3 results)
  • 2002 Annual Research Report   Final Research Report Summary
  • 2001 Annual Research Report
  • Research Products

    (50 results)

All Other

All Publications (50 results)

  • [Publications] Q.B.Tian, et al.: "Specific protein-DNA and protein-protein interaction in the hig gene system, a plasmid-borne proteic killer gene system of plasmid Rts1"Plasmid. 45. 63-74 (2001)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] K.Oana, et al.: "Molecular and epidemiological study of the first outbreak of vanB type vancomycin-resistant Enterococcus faecalis in Japan"Jpn.J.Infect.Dis.. 54. 17-22 (2001)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] M.Ohnishi, et al.: "Diversification of Escherichia coli genomes : are bacteriophages the major contributors?"Trends Microbiol.. 9. 481-485 (2001)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] I.Tatsuno, et al.: "toxB gene on pO157 of enterohaemorrhagic Escherichia coli O157 : H7 is required for full epithelial cell adherence phenotype"Infect.Immun.. 69. 6660-6669 (2001)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] T.Yamaguchi, et al.: "Complete nucleotide sequence of a Staphylococcus aureus exfoliative toxin B plasmid and identification of a novel ADP-ribosyltransferase, EDIN-C"Infect.Immun.. 69. 7760-7771 (2001)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] M.Nakano, et al.: "Association of the urease gene with enterohemorrhagic Escherichcia coli strains irrespective of their serogroups"J.Clin.Microbiol.. 39. 4541-4543 (2001)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] 大西真, 黒川顕, 林哲也: "腸管出血性大腸菌O157全ゲノム配列決定の意義"蛋白質 核酸 酵素. 46. 1862-1866 (2001)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] 大西真, 林哲也: "腸管出血性大腸菌O157:H7"臨床と微生物. 28. 751-758 (2001)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] T.Murata, et al.: "Complete nucleotide sequence of plasmid Rts1 ; implications for the evolution of large plasmid genomes"J.Bacteriol.. 184. 3194-3202 (2002)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] T.Yamaguchi, et al.: "Clonal association of Staphylococcus aureus causing bullous impetigo and the emergence of new methicillin-resistant clonal groups in Kansai district in Japan"J.Infect.Dis.. 185. 1511-1516 (2002)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] M.Ohnishi, et al.: "Genomic diversity of enterohemorrhagic Escherichia coli O157 revealed by whole genome PCR Scanning"Proc.Natl.Acad.Sci.USA. 99. 17043-17048 (2002)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] 林 哲也, 大西 真: "大腸菌O157のゲノムと出血性大腸炎"現代医療. 34. 103-108 (2002)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] 大西 真, 林 哲也: "腸管出血性大腸菌O157のゲノムの多様性"日本臨床. 60. 1077-1082 (2002)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] 林 哲也, 大西 真: "腸管出血性大腸菌最新医学の遺伝的多様性とその分子基盤"最新医学. 58. 136-141 (2003)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] 林 哲也: "I.ゲノム微生物学概諭 ゲノムからみた病原細菌"日本臨床 新世紀の感染症学(下)-ゲノム・グローバル時代の感染症アップデート-. 61. 414-422 (2003)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] 林哲也: "II.感染症の遺伝子学 大腸菌"日本臨床 新世紀の感染症学(下)-ゲノム・グローバル時代の感染症アップデート-. 61. 696-702 (2003)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] M. Ohnishi, K. Kurokawa and T. Hayashi: "Diversification of Escherichia coli genomes : are bacteriophages the major contributors?"Trends Microbiol.. 9(10). 481-485 (2001)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] I. Tatsuno, M. Horie, H. Abe, T. Miki, K. Makino, H. Shinagawa, H. Taguchi, S. Kamiya, T. Hayashi and C. Sasakawa: "toxB gene on pO157 of enterohemorrhagic Escherichia coli O157 : K7 is required for full epithelial cell adherence phenotype."Infect. Immun.. 69(11). 6660-6669 (2001)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] M. Nakano, T. Iida, M. Ohnishi, K. Kurokawa, A. Takahashi, T. Tsukamoto, T. Yasunaga, T. Hayashi and T. Honda: "Association of the urease gene with enterohemorrhagic Escherichia coli strains irrespective of their serogroups."J. Clin. Microbiol.. 39(12). 4541-4543 (2001)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Q. B. Tian, M. Ohnishi, T. Murata, K. Nakayama, Y. Terawaki and T. Hayashi: "Specific protein-DNA and protein-protein interaction in the hig gene system, a plasmid-borne proteic killer gene system of plasmid Rts1."Plasmid. 45. 63-74 (2001)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] K. Oana, Y. Kawakami, M. Ohnishi, M. Ishikawa, M. Hirota, M. Tozuka, K. Atarashi, K. Baba, K. Fujiki, M. Okazaki, T. Honda and T. Hayashi: "Molecular and epidemiological study of the first outbreak of vanB type vancomycin-resistant Enterococcus faecalis in Japan."Jpn. J. Infect. Dis.. 54. 17-22 (2001)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] T. Yamaguchi, T. Hayashi, H. Takami, M. Ohnishi, T. Murata, K. Nakayama, K. Asakawa, M. Ohara, H. Komatsuzawa and M. Sugai: "Complete nucleotide sequence of a Staphylococcus aureus exfoliative toxin B plasmid and identification of a novel ADP-ribosyltransferase, EDIN-C."Infect. Immun.. 69(12). 7760-7771 (2001)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] M. Ohnishi, K. Kurokawa and T. Hayashi: "Determination of the whole genome sequence of enterohemorrhagic Escherichia coli O157 : H7."Protein, Nucleic Acid and Enzyme. 46(12). 1862-1866 (2001)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] T. Hayashi and M. Ohnishi: "Enterohemorrhagic Escherichia coli O157 : H7."Clinical Microbiology. 28(6). 751-758 (2001)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] T. Murata, M. Ohnishi, T. Ara, J. Kaneko, C-G. Han, Y-F. Li, K. Takashima, H. Nojima, K. Nakayama, A. Kaji, Y. Kamio, T. Miki, H. Mori, E. Ohtsubo, Y. Terawaki and T. Hayashi: "Complete nucleotide sequence of plasmid Rts1 ; implications for the evolution of large plasmid genomes."J. Bacteriol.. 184(12). 3194-3202 (2002)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] T. Yamaguchi, Y. Yokota, J. Terajima, T. Hayashi, M. Aepfelbacher, M. Ohara, H. Komatsuzawa, H. Watanabe and M. Sugai: "Clonal association of Staphylococcus aureus causing bullous impetigo and the emergence of new methicillin-resistant clonal groups in Kansai district in Japan."J. Infect. Dis.. 185(10). 1511-1516 (2002)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] M. Ohnishi, J. Terajima, K. Kurokawa, K. Nakayama, Y. Ogura, H. Tamura, H. Watanabe and T. Hayashi: "Genomic diversity of enterohemorrhagic Escherichia Coli O157 revealed by whole genome PCR scanning."Proc. Natl. Acad. Sci. USA. 99(26). 17043-17048 (2002)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] T. Hayashi and M. Ohnishi: "Genome analysis of Escherichia coli O157 and its implication on hemorrhagic colitis."New Horizon for Medicine. 34(5). 103-108 (2002)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] M. Ohnishi and T. Hayashi: "Genomic diversity of enterohemorrhagic Escherichia coli O157."Japanese Journal of Clinical Medicine. 60(6). 1077-1082 (2002)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] T. Hayashi and M. Ohnishi: "Molecular basis for the genetic diversity in enterohemorrhagic Escherichia coli O157."SAISHIN IGAKU. 58(1). 136-141 (2003)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] T. Hayashi: "Genome analysis of pathogenic bacteria."Japanese Journal of Clinical Medicine. 61, Suppl. 3. 414-422 (2003)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] T. Hayashi: "Escherichia coli"Japanese Journal of Clinical Medicine. 61, Suppl. 3. 696-702 (2003)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] T.Murata, et al.: "Complete nucleotide sequence of plasmid Rts1 ; implications for the evolution of large plasmid genomes"J. Bacteriol.. 184. 3194-3202 (2002)

    • Related Report
      2002 Annual Research Report
  • [Publications] T.Yamaguchi, et al.: "Clonal association of Staphylococcus aureus causing bullous impetigo and the emergence of new methicillin-resistant clonal groups in Kansai district in Japan"J. Infect. Dis.. 185. 1511-1516 (2002)

    • Related Report
      2002 Annual Research Report
  • [Publications] M.Ohnishi, et al.: "Genomic diversity of Escherichia coil O157 revealed by whole genome PCR Scanning"Proc. Natl. Acad. SCi. USA. 99. 17043-17048 (2002)

    • Related Report
      2002 Annual Research Report
  • [Publications] 林 哲也, 大西 真: "大腸菌O157のゲノムと出血性大腸炎"現代医療. 34. 103-108 (2002)

    • Related Report
      2002 Annual Research Report
  • [Publications] 大西 真, 林 哲也: "腸管出血性大腸菌O157のゲノムの多様性"日本臨床. 60. 1077-1082 (2002)

    • Related Report
      2002 Annual Research Report
  • [Publications] 林 哲也, 大西 真: "腸管出血性大腸菌O157の遺伝的多様性とその分子基盤"最新医学. 58. 136-141 (2003)

    • Related Report
      2002 Annual Research Report
  • [Publications] 林 哲也: "I.ゲノム微生物学概論 ゲノムからみた病原細菌"日本臨床 新世紀の感染症学(下)-ゲノム・グローバル時代の感染症アップデート-. 61. 414-422 (2003)

    • Related Report
      2002 Annual Research Report
  • [Publications] 林哲也: "II. 感染症の遺伝子学 大腸菌"日本臨床 新世紀の感染症学(下)-ゲノム・グローバル時代の感染症アップデート-. 61. 696-702 (2003)

    • Related Report
      2002 Annual Research Report
  • [Publications] Q.B.Tian, et al.: "Specific protein-DNA and protein-protein interaction in the hig gene system, a plasmid-borne proteic killer gene system of plasmid Rts1"Plasmid. 45. 63-74 (2001)

    • Related Report
      2001 Annual Research Report
  • [Publications] K.Oana, et al.: "Molecular and epidemiological study of the first outbreak of vanB type vancomycin-resistant Enterococcus faecalis in Japan"Jpn.J.Infect.Dis.. 54. 17-22 (2001)

    • Related Report
      2001 Annual Research Report
  • [Publications] M.Ohnishi, et al.: "Diversification of Escherichia coli genomes : are bacteriophages the major contributors ?"Trends Microbiol.. 9. 481-485 (2001)

    • Related Report
      2001 Annual Research Report
  • [Publications] I.Tatsuno, et al.: "toxB gene on pO157 of enterohaemorrhagic Escherichia coli O157 : H7 is required for full epithelial cell adherence phenotype"Infect.Immun.. 69. 6660-6669 (2001)

    • Related Report
      2001 Annual Research Report
  • [Publications] T.Yamaguchi, et al.: "Complete nucleotide sequence of a Staphylococcus aureus exfoliative toxin B plasmid and identification of a novel ADP-ribosyltransferase, EDIN-C"Infect.Immun.. 69. 7760-7771 (2001)

    • Related Report
      2001 Annual Research Report
  • [Publications] M.Nakano, et al.: "Association of the urease gene with enterohemorrhagic Escherichcia coli strains irrespective of their serogroups"J.Clin.Microbiol.. 39. 4541-4543 (2001)

    • Related Report
      2001 Annual Research Report
  • [Publications] 大西真, 黒川顕, 林哲也: "腸管出血性大腸菌O157全ゲノム配列決定の意義"蛋白質 核酸 酵素. 46. 1862-1866 (2001)

    • Related Report
      2001 Annual Research Report
  • [Publications] 大西真, 林哲也: "腸管出血性大腸菌O157:H7"臨床と微生物. 28. 751-758 (2001)

    • Related Report
      2001 Annual Research Report
  • [Publications] 林 哲也 他: "医科細菌学第3版"南江堂. (2001)

    • Related Report
      2001 Annual Research Report
  • [Publications] 林 哲也 他: "細菌毒素ハンドブック"サイエンスフォーラム社. 587 (2002)

    • Related Report
      2001 Annual Research Report

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Published: 2001-04-01   Modified: 2016-04-21  

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