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1995 Fiscal Year Final Research Report Summary

Study of the virulence factors from periodontopathic bacteria

Research Project

Project/Area Number 06671832
Research Category

Grant-in-Aid for General Scientific Research (C)

Allocation TypeSingle-year Grants
Research Field Morphological basic dentistry
Research InstitutionTOKYO DENTAL COLLEGE

Principal Investigator

KATO Tetsuo  Tokyo Dental College, Department of Microbiology, Associate Professor, 歯学部, 助教授 (00159253)

Co-Investigator(Kenkyū-buntansha) YAMANAKA Ayumi  Tokyo Dental College, Department of Microbiology, Assistant, 歯学部, 助手 (40231667)
ISHIHARA Kazuyuki  Tokyo Dental College, Department of Microbiology, Assistant Professor, 歯学部, 講師 (00212910)
Project Period (FY) 1994 – 1995
Keywordsperiodontitis / periodontopathic bacteria / molecular cloning / endotoxin / kanamycin-resistant / hemolytic activity / virulence factor
Research Abstract

Porphyromonas gingivalis, a gram-negative anaerobic rod, has been implicated as a periodontal pathogen. Black-pigmented anaerobic rods such as P.gingivalis show especially high resistance to kanamycin (KM) as compared with the other anaerobic bacteria. A gene for KM-inactivating protein has been isolated from P.gingivalis strain 16-1 in Escheichia coli, utilizing the plasmid vector pTZ18R.The resultant kanamycin resistant E.coli clone, harboring plasmid pPG16 with a P.gingivalis DNA insert, expressed KM-inactivating activity, which was enhanced by hte addition of acetyl CoA,indicating that the KM was inactivated by acetylation. Southern blot analysis indicated that the gene is conserved among all P.gingivalis strains tested. Rabbit antiserum to P.gingivalis 16-1 whole cells reacted with a band of ca.35kDa when a sonic extract of an E.coli JM109 containing pPG16 was prepared for SDS-PAGE.These results indicate that P.gingivalis possesses KM-inactivating activity and that a gene coding for KM-inactivating activity has been isolated from P.gingivalis.
Actinobacillus actinomycetemcomitans is associated with human periodontal diseases. Recent studies have reported detectable a hemolytic activity of this organism on blood agar plates. Here we report the isolation and characterization of recombinant clone from a genomic library of A.actinomycetemcomitans ATCC 43718 that confer on E.coli the ability to lyse horse red blood cells. DNA hybridization analysis indicates that the gene was conserved among all A.actinomycetemcomitans strains tested. E.coli transformed with this gene lysed horse.sheep and human erythrocytes but not those of rabbit. Heating and treatment with proteinase K resulted in the complete hindrance of the hemolytic activity. Further study will help in understanding the function of A.actinomycetemcomitans hemolytic activity as a periodontal virulence factor.

  • Research Products

    (13 results)

All Other

All Publications (13 results)

  • [Publications] Kim,S. -J.: "B-cell mitogenicity and IL-Iβ production of Iipopolysaccharides from various Capnocytophage strains." Bull. Tokyo dont. Coll. 35. 79-83 (1994)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Hirai,K.: "The 57kDa protein is a major antigen in the outer membrane of invasive Porphyromonas gingivalis strains." Bull. Tokyo dont. Coll.35. 221-226 (1994)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Ando,T.: "Heat shock protein of periodontopathic bacteria in human poriodontitis process." Microbiol. Immunol.39. 321-327 (1995)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Harano,K.: "An antiserum to a synthetic timbrial peptide of Actinobacillus actinomycetemcomitans blocked udhesion of the microorganism." FEMS Microbiol. Lett.130. 279-286 (1995)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Ishihara,K.: "Cloning and expression of a neutral phosphutase gene from Treponema denticola." Intect. Immun.63. 1147-1152 (1995)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Kato,T.: "Identification of a chromosomally encoded kanamycin acetylase in Porphyromanas gingivalis." FEMS Microbiol. Lett.131. 301-306 (1995)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Yamanaka, A., Hirai K., Kato, T., Naito, Y., Okuda, K., Toda, S., and Okuda, K.: "Efficacy of Listerine antiseptic against MRSA,Candida albicans and HIV." Bull.Tokyo dent.Coll.35. 23-26 (1994)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Kim, S.-J., Kato, T., Naito, Y., Hirai, K., Choi, J.-I., Takazoe, I., and Okuda, K.: "B-cell mitogenicity and IL-1beta production of lipopolysaccharides from various Capnocyphaga strains." Bull.Tokyo dent.Coll.35. 79-83 (1994)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Hirai, K., Kato, T., and Okuda, K.: "The 57kDa protein is a major antigen in the outer membrane of invasive Porphyromonas gingivalis strains." Bull.Tokyo dent.Coll.35. 221-226 (1994)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Ando, T., Kato, T., Ishihara, K., Ogiuchi, H., and Okuda, K.: "Heat shock protein of periodontopathic bacteria in human periodontitis process." Microbiol.Immun.39. 321-327 (1995)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Ishihara, K., and Kuramitsu, H.K.: "Cloning and expression of a neutral phosphatase gene from Treponema denticola." Infect.Immun.63. 1147-1152 (1995)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Harano, K., Yamanaka, A.and Okuda, K.: "An antiserum to a synthetic fimbrial peptide of Actinobacillus actinomycetemcimitans blocked adhesion of the microorganism." FEMS Microbiol.Lett.130. 279-286 (1995)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Kato, T., Hirai, K.and Okuda, K.: "Identification of a chromosomally encoded kanamycin acetylase in Porphyromonas gingivalis." FEMS Microbiol.Lett.131. 301-306 (1995)

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

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Published: 1997-03-04  

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