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Molecular biological Analysis of Periodontopathic Bacteria

Research Project

Project/Area Number 01480425
Research Category

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

Allocation TypeSingle-year Grants
Research Field Morphological basic dentistry
Research InstitutionTokyo Dental College

Principal Investigator

ICHIRO Takazoe  東京歯科大学, 微生物学講座, 教授 (80085696)

Co-Investigator(Kenkyū-buntansha) YAMANAKA Ayumi  東京歯科大学, 微生物学講座, 副手
ISHIHARA Kazuyuki  東京歯科大学, 微生物学講座, 助手 (00212910)
KATO Tetsuo  東京歯科大学, 微生物学講座, 助手 (00159253)
NAITO Yuko  東京歯科大学, 微生物学講座, 講師 (00147258)
OKUDA Katsuji  東京歯科大学, 微生物学講座, 教授 (40085741)
太田 功正  東京歯科大学, 歯学部・微生物, 助教授 (30138680)
Project Period (FY) 1989 – 1991
Project Status Completed (Fiscal Year 1991)
Budget Amount *help
¥6,100,000 (Direct Cost: ¥6,100,000)
Fiscal Year 1991: ¥1,700,000 (Direct Cost: ¥1,700,000)
Fiscal Year 1990: ¥2,300,000 (Direct Cost: ¥2,300,000)
Fiscal Year 1989: ¥2,100,000 (Direct Cost: ¥2,100,000)
KeywordsPeriodontal Disease / Spirochte / Trponeme denticola / Gene cloning / ATCase / 57.5 KDa protein / Diagnosis / DNA probe / ATCase / Porphynomonas ginginalis / 表層タンパク / 付着因子 / イムノプロット / 歯周病原菌 / Bacteroides gingivalis / 遺伝子解析
Research Abstract

The mechanisms responsible for periodontal disease have not been clarified, but subgingival bacterial infection by organisms such as gram-negative rods and spirochetes is a major etiological factor in the disease process. Treponema denticola has been demonstrated to be predominant in subgingival plaque of patients with periodontitis. We have been attempting to clone and sequence several antigenic proteins of T. denticola. T. denticola ATCC 33520 was grown in TYGVS medium. Chromosomal DNA was prepared and digested with Hind III. and ligated to the phage vector L47.1. The phage particles were infected with Escherichia coli Q358. Clone of the surface antigen was pick up using rabbit antiserum against whole cells of T. denticola. The obtained clone was subcloned into plasmid vector PACYC 184 and transformed into E. coli HB101. Sonicated extracts of the cloned TD12 was approximately 57.5 KDa protein. Southern blot analysis showed that the gene exists in common in T. denticola ATCC 33521, ATCC 45404, and ATCC 35405. but does not exist in other oral treponema species. The clone protein possessed aspartate carbamoyltransferase activity and strong antigenicity. The DNA sequencing was carried out by the dideoxy chain termination method using an Applied Biosystems Model 373 automated DNA sequencer. The cloned protein contains 486 amino acids and has a calculated molecular weight of 54, 919. The cloned gene encoding for aspartate carbomyltransferase may be an useful DNA probe for identification of T. denticola and applicable for clinical examinations.

Report

(4 results)
  • 1991 Annual Research Report   Final Research Report Summary
  • 1990 Annual Research Report
  • 1989 Annual Research Report
  • Research Products

    (36 results)

All Other

All Publications (36 results)

  • [Publications] Saeki,Y.et al: "Autimicrobial action of natural substances on oral bacteria" Bull.Tokyo dent.Coll.30. 205-211 (1989)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1991 Final Research Report Summary
  • [Publications] Oosawa,K.et al: "Studies of the autibactericidal activity of plant extracts and their constituents against periodonto pathic bacteria" Bull.Tokyo dent.Coll.31. 17-21 (1990)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1991 Final Research Report Summary
  • [Publications] Kato,T.et al: "Protection of mice against the lethal toxicity of a lipopoly saccharide (LPS)by immunization with autiidiotype antibody to a monoclonal antibody to a monoclonal antibody to lipid A from Eikenella corrodens" Infect.Immun. 58. 416-420 (1990)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1991 Final Research Report Summary
  • [Publications] Oosawa,K.et al: "Inhibitony effects of aqueous extract of cacao bean husk of collagenase of Bacteroides gingivalis" Bull.Tokyo dent.Coll.31. 125-128 (1990)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1991 Final Research Report Summary
  • [Publications] Nakagawa,T.et al: "Clinical,microbiological and immunological studies following initial preparation in adult periodontitis." Bull.Tokyo dent.Coll.31. 321-331 (1990)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1991 Final Research Report Summary
  • [Publications] Ishihara,K.他: "A sensitive enzymatic method (skー013)for detection and quantification of specific pathogen" J.Periodont.Res. (1992)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1991 Final Research Report Summary
  • [Publications] Takazoe Ichiro: "Progress in Sweetners" Elsevier Applied Science, 395 (1989)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1991 Final Research Report Summary
  • [Publications] Takazoe Ichiro: "Predictable advances in oral health research" Tokyo Dental College, 190 (1991)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1991 Final Research Report Summary
  • [Publications] Kato T, Takazoe I and Okuda K: "Structural analysis of lipopolysaccharides from Eikenella corrodens by use of murine monoclonal antibodies" Infect. Immun.57. 656-659 (1989)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1991 Final Research Report Summary
  • [Publications] Saeki Y, Ito Y, Shibata M, Sato T, Okuda K and Takazoe I: "Antimicrobial action of natural substances on oral bacteria" Bull. Tokyo dent. Coll.30. 129-135 (1989)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1991 Final Research Report Summary
  • [Publications] Osawa K, Matumoto T, Maruyama T, Takiguchi T, Okuda K and Takazoe I: "Studies of the antibacterial activity of plant extracts and their constituents against periodontopathic bacteria" Bull. Tokyo dent. Coll.31. 17-21 (1990)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1991 Final Research Report Summary
  • [Publications] Kato T, Takazoe I and Okuda K: "Protection of mice against the lethal toxicity of a lipopolysacchar ide (LPS) by immunization with anti-idiotype antibody to a monoclonal antibody to lipid A from Eikenella corrodens LPS" Infect. Immun.58. 416-420 (1990)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1991 Final Research Report Summary
  • [Publications] Osawa K, Matumoto T, Maruyama T, Naito Y, Okuda K and Takazoe I: "Inhibitory effects of aqueous extract of cacao bean husk on collagenase of Bacteroides gingivalis" Bull. Tokyo dent. Coll.31. 125-128 (1990)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1991 Final Research Report Summary
  • [Publications] Choi J-I, Nakagawa T, Yamada S, Takazoe I and Okuda K: "Clinical, microbiological and immunological studies on recurrent periodontal disease" J. Clin. Periodontol.17. 426-434 (1990)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1991 Final Research Report Summary
  • [Publications] Kato T, Iijima H, Ishihara K, Kaneko T, Hirai K, Naito Y and Okuda K: "Antibacterial effects of Listerine on oral bacteria" Bull. Tokyo dent. Coll.31. 301-307 (1990)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1991 Final Research Report Summary
  • [Publications] Nakagawa T, Yamada S, Tsunoda M, Sato T, Naito Y, Takazoe I and Okuda K: "Clinical, microbiological, and immunological studies following initial preparation in adult periodontitis" Bull. Tokyo dent. Coll.31. 321-331 (1990)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1991 Final Research Report Summary
  • [Publications] Ishihara K, Naito Y, Kato T, Takazoe I and Okuda K, Eguchi T, Nakashima K, Matsuda N, Yamasaki K, Suido H, Hasegawa K and Sugihara K: "A sensitive enzymatic method (SK-013) for detection and quantification of specific periodontopathogens" J. Periodont. Res.

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1991 Final Research Report Summary
  • [Publications] Saeki,Y.et al: "Autimicrobial action of natural substances on oral bacteria" Bull.Tokyo dent.Coll.30. 205-211 (1989)

    • Related Report
      1991 Annual Research Report
  • [Publications] Oosawa,K.et al: "Studies of the autibactericidal activity of plant extracts and their constituents against periodonto pathic bacteria" Bull.Tokyo dent.Coll.31. 17-21 (1990)

    • Related Report
      1991 Annual Research Report
  • [Publications] Kato,T.et al: "Protection of mice against the lethal toxicity of a lipopolysaccharide(LPS) by immunization with autiidiotype antibody to a monodonal antibody to a monodclonal antibody to lipid A from Eikenella corrodens" Infect.Immun. 58. 416-420 (1990)

    • Related Report
      1991 Annual Research Report
  • [Publications] Oosawa,K.et al: "Inhibitony effects of aqueous extract of cacao bean husk of collagenase of Bacteroides gingivalis" Bull.Tokyo dent.Coll.31. 125-128 (1990)

    • Related Report
      1991 Annual Research Report
  • [Publications] Nakagawa,T.et al: "Clinical,microbiological and immunological studies following initial preparation in adult periodontitis." Bull.Tokyo dent.Coll.31. 321-331 (1990)

    • Related Report
      1991 Annual Research Report
  • [Publications] Ishihara,K.他: "A sensitive enzymstic method (skーo13)for detection and quantification of specific pathogen" J.Periodont.Res.(1992)

    • Related Report
      1991 Annual Research Report
  • [Publications] Takazoe Ichiro: "Progress in Sweetners" Elsevier Applied Science, 395 (1989)

    • Related Report
      1991 Annual Research Report
  • [Publications] Takazoe Ichiro: "Predictable advances in oral health research" Tokyo Dental College, 190 (1991)

    • Related Report
      1991 Annual Research Report
  • [Publications] 高添 一郎: "口腔常在菌と感染" メディカルコ-ナ-. 84. 12-14 (1989)

    • Related Report
      1990 Annual Research Report
  • [Publications] 高添 一郎: "低齲蝕甘味料開発の動向" 医療. 89. 66-68 (1989)

    • Related Report
      1990 Annual Research Report
  • [Publications] Saeki,Y.,Ito,Y.,Shibata,M.,Sato,Y.,Okuda,K.and Takazoe,I.: "Antimicrobial action of natural substances on bacteria" Bull.Tokyo Dental College. 30. 129-135 (1989)

    • Related Report
      1990 Annual Research Report
  • [Publications] Osawa,K.,Matsumoto,T.,Maruyama,T.,Takiguch,T.,Okuda,K.and Takazoe,I.: "Studies of the antibacterial activity of plant extracts and their constituents against periodontopathic bacteria" Bull.Tokyo Dental College. 31. 17-21 (1990)

    • Related Report
      1990 Annual Research Report
  • [Publications] Kato.T.,Takazoe,I.,Okuda,K.: "Protection of mice against the lethal toxicity of a lipopolyーsaccharide (LPS) by immunization with antiーidiotype antibody to a monoclonal antibody to lipid A from Eikenella corrodens" Infection and Immunity. 58. 416-420 (1990)

    • Related Report
      1990 Annual Research Report
  • [Publications] Choi.J.I.,Nakagawa.T.,Yamad,S.,Takazoe,I.,Okuda,K.: "Clinical,Microbiological and immunological studies on vecurrent periodontal disease" Journal of Clinical Periodontology. 17. 426-434 (1990)

    • Related Report
      1990 Annual Research Report
  • [Publications] Takazoe,I.著分担: "Progress in Sweetners" Elsevier Applied Science,London,

    • Related Report
      1990 Annual Research Report
  • [Publications] 高添 一郎著分担: "標準微生物学" 医学大院, 463 (1990)

    • Related Report
      1990 Annual Research Report
  • [Publications] Kato,T.: "Structural analysis of lipo-polysaccharides from Eikenella corrodens by use of murine monoclonal antibodies." Infect.Immun.57. 656-659 (1989)

    • Related Report
      1989 Annual Research Report
  • [Publications] Fukumoto,Y.: "A common antigen of Treponema denticola and other Treponema species detected by monoclonal antibody." Oral Microbiol Immunol. 4. 112-116 (1989)

    • Related Report
      1989 Annual Research Report
  • [Publications] Okuda,K.: "Neuraminidase-enhanced attachment of Bacteroides intermedius to human erythrocytes and buccal epithelial cells." Infect.Immun.57. 1635-1637 (1989)

    • Related Report
      1989 Annual Research Report

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Published: 1989-04-01   Modified: 2016-04-21  

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