| Project/Area Number |
12470464
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| Research Category |
Grant-in-Aid for Scientific Research (B)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
矯正・小児・社会系歯学
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| Research Institution | Kagoshima University |
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Principal Investigator |
INOUE Masakazu Kagoshima University, Dental School, Professor, 歯学部, 教授 (30028740)
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| Co-Investigator(Kenkyū-buntansha) |
ITO Hiro-o Kagoshima University, Dental School, Associate Professor, 歯学部, 助教授 (40213079)
SATO Setsuko Kagoshima University, Dental Hospital, Assistant Professor, 歯学部附属病院, 講師 (70145514)
KITADA Katsuhiro Kagoshima University, Dental School, Research Associate, 歯学部, 助手 (90195264)
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| Project Period (FY) |
2000 – 2002
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| Project Status |
Completed (Fiscal Year 2002)
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| Budget Amount *help |
¥12,000,000 (Direct Cost: ¥12,000,000)
Fiscal Year 2002: ¥1,500,000 (Direct Cost: ¥1,500,000)
Fiscal Year 2001: ¥2,700,000 (Direct Cost: ¥2,700,000)
Fiscal Year 2000: ¥7,800,000 (Direct Cost: ¥7,800,000)
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| Keywords | Nutritionally variant streptococci (NVS) / Granulicatella adiacens / Oral indigenous bacteria / Infective endocarditis / Fibronectin / Gene cloning / 2', 3' -cyclic nucleotide 2' -phosphodiesterase / Polymerase chain reaction (PCR) / 細胞外基質タンパク質 / DNAライブラリー / 発現ライブラリー / 蛍光標識 |
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| Research Abstract |
A gene encoding a protein with an ability of fibronectin binding was cloned from Granulicatella adiacens ATCC 49175T. DNA sequence revealed that the gene was located from 806 to 2419 on the 2540-bp EcoRI-XbaI fragment from the genomic DNA of G adiacens. The deduced amino acid sequence showed 54% identity to Lactococcus lactis 2', 3' -cyclic nucleotide 2' -phosphodiesterase (EC 3.1.4.16). An antibody specific to the recombinant protein was prepared in mice, but it was not able to bind to the cell surface of G adiacens, suggesting that the protein is not exposed on the surface of the organism. A set of PCR primers was designed according to the DNA sequences of the gene and DNA from various species were subjected to amplification. A 0.52-kb PCR product was yielded only from strains of G adiacens and closely related species produce no product. Clinical specimens of dental plaques from healthy volunteers produced a single 0.52-kb product, as did the type strain G adiacens. These results suggest that the PCR primers are applicable to detection and identification of G adiacens for clinical diagnosis.
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