| Project/Area Number |
11660284
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Applied animal science
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| Research Institution | KOKKAIDO UNIVERSITY (2000)
Mie University (1999) |
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Principal Investigator |
KOBAYASHI Yasuo Hokkaido Univ. Grad.School of Agr., Asso.Prof., 大学院・農学研究科, 助教授 (50153648)
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| Project Period (FY) |
1999 – 2000
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| Project Status |
Completed (Fiscal Year 2000)
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| Budget Amount *help |
¥2,800,000 (Direct Cost: ¥2,800,000)
Fiscal Year 2000: ¥800,000 (Direct Cost: ¥800,000)
Fiscal Year 1999: ¥2,000,000 (Direct Cost: ¥2,000,000)
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| Keywords | Rumen / Cellulolytic bacteria / Competitive PCR / Sheep / DNA / DNA抽出 / 定量 / 特異プライマー / 付着 |
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| Research Abstract |
The present study was attempted to analyze ecology of three representative cellulolytic bacterial species in the rumen by newly established molecular biological techniques that were competitive PCR and FISH.The results obtained througt development of the techniques and their application are follows.
1. Primers designed by using 16SrDNA sequences specific to the target bacterial species allowed specific amplification of the corresponding species. The developed cPCR assays were highly quantitative at pure and mixed culture levels, then the assay values linearly responded to the addition of known amount of target bacteria. The assay values were satisfactorily reproducible.
2. In application of the assays, it was revealed that F.succinogenes wa more dominant in sheep rumen than two ruminococcal species, especially when a high roughage diet was given. F.succinogenes was quantitated in the hindgut at a level similar to that in the rumen. These are indicative of major contribution of this bacterial species to cellulolysis in the gastrointestinal tract.
3. FISH was developed to visualize the target bacterial species under microscopy. After condition of hybridization of the bacterial RNA with a fluorescence-labelled DNA probe was established, the target bacteria was detected at pure and mixed culture levels.
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