| Project/Area Number |
03454067
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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 |
応用微生物学・発酵学
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| Research Institution | Nagaoka University of technology |
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Principal Investigator |
YANO Keiji Nagaoka Univ. Tech., Dept. BioEngineering, Professor, 工学部, 教授 (10011842)
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| Co-Investigator(Kenkyū-buntansha) |
KIMBARA Kazuhide Nagaoka Univ. Tech., Dept. BioEngineering, Research Associate, 工学部, 助手 (30225122)
FUKUDA Masao Nagaoka Univ. Tech., Dept. BioEngineering, Associate Professor, 工学部, 助教授 (20134512)
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| Project Period (FY) |
1991 – 1992
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| Project Status |
Completed (Fiscal Year 1992)
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| Budget Amount *help |
¥6,600,000 (Direct Cost: ¥6,600,000)
Fiscal Year 1992: ¥1,100,000 (Direct Cost: ¥1,100,000)
Fiscal Year 1991: ¥5,500,000 (Direct Cost: ¥5,500,000)
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| Keywords | PCB / Degradation Genes / Cloning / PCR / Molecular Evolution / クロ-ニング |
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| Research Abstract |
In these experiment, we analyzed the structure of PCB degradation genes and discussed about their evolutionary origin. The gene responsible for 2,3-dihydroxybiphenyl dioxygenase (bphc) was screened from several bacteria and were cloned for further analysis. A PCR method was used for screening of biphenyl degradation genes. However this method was suitable only for highly homologous genes.
Recently we have isolated a bacterium from soil which can degrade not only PCB but also benzene and toluene. This bacterium can degrade all of the PCB congeners. It is a gram positive bacterium which was identified as a species of Rhodococcus. Degradative genes were cloned and the sequencing of bphC was done. A computer analysis of homology of bphC related genes was done and a phylogenic tree was constructed. These results suggested that amino acid differences effect the differences of the activity.
To monitor genetically manipulated microorganism in the environment, we have devised a method for isolating total bacterial DNA from soil. Bacterial fraction was concentrated by ultracentrifuge and stored in deep freezer. Total DNA was isolated from this bacterial fraction. DNA hybridization was done with the soil isolated DNA using biphenyl degradation genes as probes, but a positive signal was not observed. The hybridization conditions including probe designing should be changed for a better understanding.
We conclude from our experiment that the evolution of degradative genes are effected by the environment such as the living condition of these bacteria.
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