| Project/Area Number |
05671515
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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 | Showa University |
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Principal Investigator |
YAMAMOTO Ayako Showa University, school of Dentistry, Associated Prof., 歯学部, 助教授 (20085773)
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| Co-Investigator(Kenkyū-buntansha) |
GOTO Nobuichi Showa University, school of Dentistry, Prof., 歯学部, 教授 (10077175)
菊田 典子 昭和大学, 歯学部, 助手 (50255874)
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| Project Period (FY) |
1993 – 1994
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| Project Status |
Completed (Fiscal Year 1994)
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| Budget Amount *help |
¥2,000,000 (Direct Cost: ¥2,000,000)
Fiscal Year 1994: ¥500,000 (Direct Cost: ¥500,000)
Fiscal Year 1993: ¥1,500,000 (Direct Cost: ¥1,500,000)
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| Keywords | Entamoeba gingivalis / Oral amoeba / SrRNA / Species-specific DNA probe / Phylogenetic analysis / Protozoan / Phylogenetic analysis / ribosomal DNA probe / Oligomucleotide probe / digoxigenin labeled probe / oral amoeba / in situ hybridization |
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| Research Abstract |
The small subunit ribosomal RNA (SrRNA) gene of Entamoeba gingivalis was amplified by PCR and the product of 1.9-kb long was cloned into a plasmid vector pUC18. Four clones were isolated and sequenced. The insert DNAs were 1918-to 1921-bp long and A+T rich (65.5%). The four SrRNA sequences of E.gingivalis were aligned with those of nine related protozoans searching for E.gingivalis-specific sequence. A sequence of 28 oligonucleotides was chosen, chemically synthesized, and labeled with digoxigenin for a DNA probe. The probe thus constructed was shown to hybridize only with either the SrRNA-coding DNAs or the cells of the two E.gingivalis strains and not with those of other protozoans and oral fungi tested. A representative SrRNA-sequence was analyzed and a phylogenetic tree was constructed by neighbor-joining (NJ) method. Among the protists examined, E.gingivalis was placed next to Entamoeba histolytica as expected from the traditional taxonomy.
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