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Assembly of alternating polymerized mediator, polymerized coenzyme, and functional nano-device by layer-by-layer adsorption technique

Research Project

Project/Area Number 16510084
Research Category

Grant-in-Aid for Scientific Research (C)

Allocation TypeSingle-year Grants
Section一般
Research Field Nanomaterials/Nanobioscience
Research InstitutionUniversity of Fukui

Principal Investigator

SUYE Shin-ichiro  University of Fukui, Applied Chemistry and Biotechnology, Associate Professor, 工学研究科, 助教授 (90206376)

Project Period (FY) 2004 – 2005
Project Status Completed (Fiscal Year 2005)
Budget Amount *help
¥3,000,000 (Direct Cost: ¥3,000,000)
Fiscal Year 2005: ¥1,300,000 (Direct Cost: ¥1,300,000)
Fiscal Year 2004: ¥1,700,000 (Direct Cost: ¥1,700,000)
KeywordsDNA sensor / Salmonella / Layer-by-layer / Polymerized mediator / Electron transfer / Nano-device / 分子配向 / サンドイッチハイブリダイゼーション
Research Abstract

Nucleic acid hybridization is a basic method in molecular biology and provides new possibilities in various biomedical and biochemical areas. Conventionally, fluorescence compound labeled polynucleotides were used for probes for detection of complimentary DNA by specific hybridization. However, fluorescence detection has been limited to clinical diagnosis by cost problems. In the present study, a DNA electrochemical sensor system is proposed based on the combination of sandwich hybridization of reporter probe, capture probe, and target DNA. A biotinylated DNA probe was used for capture probe for target DNA on the avidin immobilized electrode. InvA gene of Salmonella typhimurium was used for target DNA and Glucose-6-phosphate dehydrogenase was used for subsequent enzymatic electrochemical detection as reporter probe. Origonucleotide for capture probe : 5'-CACCAATATCGCCAGTACGA-biotinylated-3' and reporter probe : 5'-biotinylated-GCAGCATTAGGTATCCTG-3' were designed. Biotinylated Glucose-6 … More -dehydrogenase (G6PDH, Unitika Co.) was immobilized on the 5' end of reporter probe via avidine. DNA (invA region) was prepared form cell culture of S.typhimurium by PCR. DNA sensor was constructed as follows. At first, Au electrode was modified with mercaptopropionic acid. Then PEI-Fc (ferrocene immobilized polyethylenimine)/alginic acid and diahorase (DI)/PEI layer was formed on the surface of electrode by layer-by-layer method. Finally capture probe was immobilized on the electrode via avidine. Hybridization of target DNA and the both probe on the electrode was carried at 96□. After hybridization with target DNA, electrochemical measurement was performed in the solution containing G6P as substrate and NAD+ as cofactor for enzyme reaction. As shown in Fig.2, the anodic current (at ca. 0.25 V vs. Ag/AgCl) of ferrocene moieties of PEI-Fc raised. It indicates that reporter probe was immobilized on the electrode by hybridization with target DNA and G6PDH on the probe produced NADH. The proposed method was applicable to the measurement of DNA for environmental analysis, food analysis, and etc. Less

Report

(3 results)
  • 2005 Annual Research Report   Final Research Report Summary
  • 2004 Annual Research Report
  • Research Products

    (22 results)

All 2005 2004 Other

All Journal Article (19 results) Book (2 results) Patent(Industrial Property Rights) (1 results)

  • [Journal Article] Assembly of alternating polymerized mediator,polymerized coenzyme, and enzyme modified electrode by layer-by-layer adsorption technique2005

    • Author(s)
      S.Suye, H.Zheng, H.Okada, T.Hori
    • Journal Title

      Sensor and Actuator B 108

      Pages: 671-675

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2005 Final Research Report Summary
  • [Journal Article] Amperometric DNA sensor using gold electrode modified with polymeric mediator by layer-by-layer adsorption2005

    • Author(s)
      H.Zheng, Y.Hirose, T.Kimura, S.Suye, T.Hori, H.Katayama, J.Arai
    • Journal Title

      Microelectronics Engineering 81

      Pages: 441-447

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2005 Final Research Report Summary
  • [Journal Article] レポーター・キャプチャープローブを用いた交互積層電極によるDNAセンシングシステム2005

    • Author(s)
      末 信一朗
    • Journal Title

      ジャパンフードサイエンス 44

      Pages: 23-26

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2005 Annual Research Report 2005 Final Research Report Summary
  • [Journal Article] Assembly of alternating polymerized mediator, polymerized coenzyme, and enzyme modified electrode by layer-by-layer adsorption technique2005

    • Author(s)
      S.Suye, H.Zheng, H.Okada, T.Hori
    • Journal Title

      Sensor and Actuator B 108

      Pages: 671-675

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2005 Annual Research Report 2005 Final Research Report Summary
  • [Journal Article] DNA sensing system using modified electrode with capture and reporter probe by layer-by-layer adsorption2005

    • Author(s)
      S.Suye
    • Journal Title

      Japan Food Science (in Japanese) 44

      Pages: 23-26

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2005 Final Research Report Summary
  • [Journal Article] Amperometric DNA sensor using gold electrode modified with polymeric mediator by layer-by-layer adsorption2005

    • Journal Title

      Microelectronics Engineering 81

      Pages: 441-441

    • Related Report
      2005 Annual Research Report
  • [Journal Article] Layer-by-layer assembly of enzymes and polymerized mediator on electrode surface by electrostatic adsorption2004

    • Author(s)
      H.Zheng, H.Okada, S.Nojima, S.Suye, T.Hori
    • Journal Title

      Sci.Technol.Adv.Mat., 5

      Pages: 371-376

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2005 Final Research Report Summary
  • [Journal Article] Autonomous multielectrode system for monitoring the interactions of isoflavonoids with lung cancer cells2004

    • Author(s)
      S.Andreescu, O.A.Sadik, D.W.McGee, S.Suye
    • Journal Title

      Analtical Chemistry 76

      Pages: 2321-2330

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2005 Final Research Report Summary
  • [Journal Article] A tubulin-based quantitative assay for Taxol (paclitaxel) with enzyme channeling sensing2004

    • Author(s)
      S.Suye, D.Nakamoto, T.Hori, F.Sugawara, A.Mulchandani
    • Journal Title

      Electroanalysis 18

      Pages: 688-690

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2005 Final Research Report Summary 2004 Annual Research Report
  • [Journal Article] Amperometric DNA sensor using gold electrode modified with polymeric mediator by layer-by-layer adsorption2004

    • Author(s)
      S.Suye, T.Kimura, H.Zheng, T.Hori, Y.Amano, H.Katayama
    • Journal Title

      Chemical Sensor Supplement B 20

      Pages: 512-513

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2005 Final Research Report Summary 2004 Annual Research Report
  • [Journal Article] Layer-by-layer assembly of enzymes and polymerized mediator on electrode surface by electrostatic adsorption2004

    • Author(s)
      H.Zheng, H.Okada, S.Nojima, S.Suye, T.Hori
    • Journal Title

      Sci.Technol.Adv.Mat. 5

      Pages: 371-376

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2005 Final Research Report Summary
  • [Journal Article] Layer-by-layer assembly of enzymes and polymerized mediator on electrode surface by electrostatic adsorption2004

    • Author(s)
      H.Zheng, H.Okada, S.Nojima, S.Suye, T.Hori
    • Journal Title

      Science and Technology of Advanced Materials 5

      Pages: 371-376

    • Related Report
      2004 Annual Research Report
  • [Journal Article] Autonomous multielectrode system for monitoring the interactions of isoflabonoids with lung cancer cells2004

    • Author(s)
      S.Andreescu, O.A.Sadik, D.W.McGee, S.Suye
    • Journal Title

      Analtical Chemistry 76

      Pages: 2321-2330

    • Related Report
      2004 Annual Research Report
  • [Journal Article] L-Proline sensor based on layer-by-layer immobilization of theromstable dye-linked L-proline dehydrogenase and polymerized mediator

    • Author(s)
      H.Zheng, Y.Hirose, T.Kimura, S.Suye, T.Hori, H.Katayama, J.Arai, R.Kawakami, T.Ohshima
    • Journal Title

      Sci.Technol.Adv.Mat., (印刷中)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2005 Final Research Report Summary
  • [Journal Article] L-Proline sensor based on layer-by-layer immobilization of theromstable dye-linked L-proline dehydrogenase and polymerized mediator

    • Author(s)
      H.Zheng, Y.Hirose, T.Kimura, S.Suye, T.Hori, H.Katayama, J.Arai, R.Kawakami, T.Ohshima
    • Journal Title

      Sci.Technol.Adv.Mat. (in press)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2005 Final Research Report Summary
  • [Journal Article] Amperometric DNA sensor using gold electrode modified with polymeric mediator by layer-by-layer adsorption

    • Author(s)
      S.Suye, H.Zheng
    • Journal Title

      Smart Biosensor (G.K.Knopf, ed.) (Marcell Deckker, NY) (in press)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2005 Final Research Report Summary
  • [Journal Article] L-Proline sensor based on layer-by-layer immobilization of theroms table dye-linked L-proline dehydrogenase and polymerized mediator

    • Author(s)
      H.Zheng, Y.Hirose, T.Kimura, S.Suye, T.Hori, H.Katayama, J.Arai, R.Kawakami, T.Ohshima
    • Journal Title

      Sci.Technol.Adv.Mat., (印刷中)

    • Related Report
      2005 Annual Research Report
  • [Journal Article] Assembly of alternating polymerized mediator, polymerized coenzyme, and enzyme modified electrode by layer-by-layer adsorption technique

    • Author(s)
      S.Suye, H.Zheng, H.Okada, T.Hori
    • Journal Title

      Sensor and Actuator B (印刷中)

    • Related Report
      2004 Annual Research Report
  • [Journal Article] Amperometric DNA sensor using gold electrode modifies with polymeric mediator by layer-by-layer adsorption

    • Author(s)
      S.Suye, T.Matsuura, T.Kimura, H.Zheng, T.Hori, Y.Amano, H.Katayama
    • Journal Title

      Microelectronics Engineering (印刷中)

    • Related Report
      2004 Annual Research Report
  • [Book] Smart Biosensor

    • Author(s)
      S.Suye, H.Zheng
    • Publisher
      G.K.Knopf,ed., Marcell Deckker,NY(印刷中)
    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2005 Final Research Report Summary
  • [Book] Smart Biosensor (G.K.Knopf.ed.)

    • Author(s)
      S.Suye, H.Zheng
    • Publisher
      Marcell Deckker, NY(印刷中)
    • Related Report
      2005 Annual Research Report
  • [Patent(Industrial Property Rights)] DNA測定装置2004

    • Inventor(s)
      末 信一朗
    • Industrial Property Rights Holder
      ダイキン工業
    • Industrial Property Number
      2004-275817
    • Filing Date
      2004-09-22
    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2005 Final Research Report Summary 2004 Annual Research Report

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Published: 2004-04-01   Modified: 2016-04-21  

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