| Project/Area Number |
05660092
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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 |
応用微生物学・応用生物化学
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| Research Institution | KYOTO UNIVERSITY |
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Principal Investigator |
KUWAHARA Masaaki Kyoto Univ. Wood Research Institute Professor, 木質科学研究所, 教授 (40035978)
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| Co-Investigator(Kenkyū-buntansha) |
WATANABE Takashi Kyoto Univ. Wood Research Institute Assoc. Prof., 木質科学研究所, 助教授 (80201200)
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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 | Basidiomycete / Host-vector system / Peroxidase / Mitochondria / Plasmid / Mn (II) -peroxidase / Pleurotus ostreatus / ペルオキシダーゼ / マンガンペルオキシダーゼ / Pleurotus ostreatus / リグニンペルオキシダーゼ / Bjerkandera adusta |
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| Research Abstract |
This research aims to isolate the marker gene which are indispensable for the construction of host-veetor system of basidiomycetes.
DNA flagments which were thought to correspond to the Mn (II) peroxidase (MnP) gene were obtained from genomic DNA of Pleurotus ostreatus by PCR.These flagments were further used as the probes to isolate the DNA of MnP gene by Southern hybridization from the genomic DNA of P.ostreatus. Nucleotide sequence obtained from the clones by the hybridization were detemined. Secondary screening was carried out on the clones obtained. Transcription of mRNA isolated from the mycelia of P.ostreatus just before the activity of MnP was certificated by using RT-PCR technique.
Two lenear mitochondrial plasmids (PO-1 and-2) were isolated from the mycelia of P.ostreatus. About 55 % regeon of the hole DNA was cloned in to the vectors of Esherichia coli.
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