| Project/Area Number |
06671832
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| Research Category |
Grant-in-Aid for General Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Research Field |
Morphological basic dentistry
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| Research Institution | TOKYO DENTAL COLLEGE |
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Principal Investigator |
KATO Tetsuo Tokyo Dental College, Department of Microbiology, Associate Professor, 歯学部, 助教授 (00159253)
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| 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)
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| Project Period (FY) |
1994 – 1995
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| Project Status |
Completed (Fiscal Year 1995)
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| Budget Amount *help |
¥2,100,000 (Direct Cost: ¥2,100,000)
Fiscal Year 1995: ¥1,000,000 (Direct Cost: ¥1,000,000)
Fiscal Year 1994: ¥1,100,000 (Direct Cost: ¥1,100,000)
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| Keywords | periodontitis / periodontopathic bacteria / molecular cloning / endotoxin / kanamycin-resistant / hemolytic activity / virulence factor |
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| 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.
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