Isolation of dechlorinating genes and enzymes from soil of the long-term experimental field where organochlorine pesticides have been applied for more than thirty years
| Project/Area Number |
16380047
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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 |
Plant nutrition/Soil science
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| Research Institution | The University of Tokyo |
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Principal Investigator |
SENOO Keishi The University of Tokyo, Graduate School of Agricultural and Life Sciences, Professor (40206652)
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| Co-Investigator(Kenkyū-buntansha) |
OTSUKA Shigeto The University of Tokyo, Graduate School of Agricultural and Life Sciences, Lecturer (10313074)
西山 雅也 東京大学, 大学院・農学生命科学研究科, 助教授 (50263801)
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| Project Period (FY) |
2004 – 2006
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| Project Status |
Completed (Fiscal Year 2006)
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| Budget Amount *help |
¥12,100,000 (Direct Cost: ¥12,100,000)
Fiscal Year 2006: ¥1,800,000 (Direct Cost: ¥1,800,000)
Fiscal Year 2005: ¥1,500,000 (Direct Cost: ¥1,500,000)
Fiscal Year 2004: ¥8,800,000 (Direct Cost: ¥8,800,000)
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| Keywords | soil / long-term experimental field / γ-HCH / soil DNA / lin genes / dioxygenase genes / soil microoraanisms / ecosystem restoration / IS6100 / 石油成分 / ジオキシゲナーゼ / γ-BHC / 脱塩素遺伝子 / lin遺伝子群 / γ-HCH分解菌 / Sphingomonas paucimobilis SS86 / Sphingomonas sp. / linA / linB / ハロアルカン脱ハロゲン酵素 |
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| Research Abstract |
1. Isolation of IinB (haloalkane dehalogenase) with sequence diversity
Five strains of novel γ-HCH-degrading Sphingbium were obtained without enrichment culture. PCR amplification of genomic DNA of these strains targeting lin genes followed by cloning and sequencing were carried out. As a result, five different linB with nucleotide sequence diversity were obtained.
PCR amplification of soil DNA extracted from γ-HCH-added manure plot soil targeting genes flanked by IS6100 was carried out. Cloning and sequencing of the amplified product revealed that all of the analysed /IS6100-flanked genes were linB with sequence diversity.
2. Isolation of linA (dehaydrochlorinase) with sequence diversity
linA-targeting PCR amplification of soil DNA extracted from different plots in the long-term experimental field was carried out. The amplified products were obtained for all of the soil DNA. Cloning and sequencing of the DNA generated several linA-homologous DNA sequences with several base substitutions.
3. Isolation of petroleum-degrading genes with sequence diversity
Soils collected from different plots (manure, TPN/manure, TPN/HCH/manure) in the long-term experimental field were added with petroleum component(naphthalene, fluorine and petroleum ether) and incubated. Soil DNA extracted from the incubated soils were subjected to PCR trying to amplify naphthalene degrading genes. After cloning and sequencing analysis of the amplified products, dioxygenase gene-like DNA sequences were obtained. Not only DNA sequences highly homologous to that of known dioxygenase, but also a number of novel sequences with low homology were obtained.
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Report
(4 results)
Research Products
(3 results)
