| Project/Area Number |
61480395
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| Research Category |
Grant-in-Aid for General Scientific Research (B)
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| Allocation Type | Single-year Grants |
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| Research Field |
Functional basic dentistry
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| Research Institution | Osaka University |
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Principal Investigator |
HAMADA Shigeyuki Osaka University Faculty of Dentistry Professor, 歯学部, 教授 (60028777)
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| Co-Investigator(Kenkyū-buntansha) |
KOGA Toshihiko National Institute of Health, Dept of Dental Research, 歯科衛生部, 部長 (10037541)
OKAHASHI Nobuo National Institute of Health, Dept of Dental Research, 歯科衛生部, 研究員 (40150180)
高田 春比古 大阪大学, 歯学部, 講師 (30135743)
辻本 雅哉 大阪大学, 歯学部, 助手 (50144499)
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| Project Period (FY) |
1986 – 1988
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| Project Status |
Completed (Fiscal Year 1988)
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| Budget Amount *help |
¥3,000,000 (Direct Cost: ¥3,000,000)
Fiscal Year 1988: ¥1,500,000 (Direct Cost: ¥1,500,000)
Fiscal Year 1987: ¥1,500,000 (Direct Cost: ¥1,500,000)
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| Keywords | S / mutans / Protein antigen / Dental caries / Gene cloning / S、mutans / S, mutans / S.sanguis / DNA / プラスミド / IgAプロテアーゼ / バクテリオシン |
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| Research Abstract |
A 190 KD protein located on the cell surface of serotype c streptococcus mutans plays an important role for adherence of the organism to the tooth surface. The structural gene for the 190 KD protein antigen (PAc) of S.mutans MT8148 (type c) was cloned into the plasmid vector pUC118. SDS-polyacrylamide gel electrophoresis and western blotting showed that the E. coli harboring the chimeric plasmid produced multiple polypeptides of 190 to 210 KD. Immunodiffusion analysis revealed that the cloned PAc had the same specific determinants as the S. mutans PAc. The cloned pac gene was mapped, and it transcriptional orientation was determined by characterizing deletion mutants of the chimeric plasmid. Southern blot analysis with the cloned gene sequence as a probe revealed the presence of a homologous sequence in DNAs from serotypes c/e/f S. mutans, but not from serotypes d/g S. sobrinus. PAc-defective mutants were constructed by the insertion of an erythromycin-resistance gene into the pac gene. Cell surface hydrophobicity of the mutants was lower than that of the parent strain MT8148.
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