| Project/Area Number |
19780240
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| Research Category |
Grant-in-Aid for Young Scientists (B)
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| Allocation Type | Single-year Grants |
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| Research Field |
Boundary agriculture
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| Research Institution | Nagaoka National College of Technology |
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Principal Investigator |
TASAKI Yuji Nagaoka National College of Technology, 物質工学科, 准教授 (90390434)
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| Project Period (FY) |
2007 – 2009
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| Project Status |
Completed (Fiscal Year 2009)
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| Budget Amount *help |
¥2,820,000 (Direct Cost: ¥2,400,000、Indirect Cost: ¥420,000)
Fiscal Year 2009: ¥1,040,000 (Direct Cost: ¥800,000、Indirect Cost: ¥240,000)
Fiscal Year 2008: ¥780,000 (Direct Cost: ¥600,000、Indirect Cost: ¥180,000)
Fiscal Year 2007: ¥1,000,000 (Direct Cost: ¥1,000,000)
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| Keywords | 環境 / 水資源 / 遺伝子 / 応用微生物 / 酵素 / バイオテクノロジー / 農薬補助剤 / アルキルフェノールポリエトキシレート / 微生物分解 / 内分泌かく乱物質 / トランスポゾンタギング / Pseudomonas属細菌 |
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| Research Abstract |
Alkylphenol polyethoxylates (APEOs) belonging to a group of nonionic surfactants are biodegraded by bacteria and then short ethoxy chain-alkylphenols which are potential endocrine disrupters are produced. In this study, to elucidate the biodegradation mechanism of APEOs, we isolated genes that were involved in biodegradation of octylphenol polyethoxylate (OPEO), a kind of APEOs, from OPEO degrader Pseudomonas putida S-5 by transposon tagging and analyzed their DNA sequences. Approximately 11,000 transposon mutants were constructed by insertion of transposon Tn5 into P.putida S-5. Seventeen mutants that are not able to metabolize OPEO, namely OPEO-degradation gene disruptants, were isolated. DNA sequences of the genes disrupted by insertion of transposon were determined. A similarity search showed that 17 proteins encoded by the genes have high amino acid sequence similarity to acyl-CoA synthetase, chaperone protein, NADH quinone oxidoreductase, etc. Gene expression of 9 genes in P.putida S-5 cells grown on different carbon source (OPEO or glucose) was analyzed by RT-PCR. T424 gene was expressed only in cells grown on OPEO, while the other genes were expressed in cells grown on glucose as well as on OPEO.
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