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Development of Bioaffinity Sensor for DNA Mutation Assay

Research Project

Project/Area Number 09555266
Research Category

Grant-in-Aid for Scientific Research (B)

Allocation TypeSingle-year Grants
Section展開研究
Research Field 工業分析化学
Research InstitutionKYUSHU UNIVERSITY

Principal Investigator

MAEDA Mizuo  Kyushu University, Materials Physics and Chemistry, Professor, 工学部, 教授 (10165657)

Co-Investigator(Kenkyū-buntansha) KATAYAMA Yoshiki  Kyushu University, Materials Physics and Chemistry, Associate Professor, 工学部, 助教授 (70284528)
Project Period (FY) 1997 – 1998
Project Status Completed (Fiscal Year 1998)
Budget Amount *help
¥6,300,000 (Direct Cost: ¥6,300,000)
Fiscal Year 1998: ¥2,800,000 (Direct Cost: ¥2,800,000)
Fiscal Year 1997: ¥3,500,000 (Direct Cost: ¥3,500,000)
KeywordsDNA biosensor / DNA / gene assay / gene mutation / gene sensor / cyclic voltammetry / DNA binding protein / gene
Research Abstract

Most of DNA biosensors direct toward the detection of target DNA which has a complementary base sequence of the immobilized DNA on the electrode. On the other hand, DNA molecule can be regarded as ligands to various DNA binding proteins which recognize certain base sequences, mismatch base pair, or characteristic structure of DNA.We investigated the possibility of use of DNA binding proteins to the design of biosensor for gene assay. First, we established the system, in which DNA-protein interaction can be detected, using anti-DNA antibody and DNA modified gold electrode. In this system, ferrocyanide/ferricyanide redox couple was used as anionic redox marker to monitor the protein binding on immobilized DNA.Thus, addition of anti-DNA antibody suppressed the redox currents of marker ions dramatically in concentration dependent manner with the detection limit of 1 nM.In this case, 2-mercaptoethanol treatment of electrode surface was effective to avoid non-specific adsorption of other proteins. Then, DNA detection with the anti-DNA antibody immobilized electrode was also investigated in detail. After the study using the antibody, DNA binding protein was changed to PIT-1, which is a transcriptional factor, as sequence specific DNA binding protein. The redox current of the anionic redox marker was then augmented with the addition of PIT-i in the DNA immobilized electrode system, probably due to the cationic charges of PIT-1 which accelerates the electrochemical reaction of the marker ions. The increase of the redox current was again the concentration dependent of added PIT-1.
In this way, we successfully established the detecting system of sequence specific interaction of DNA-DNA binding protein.

Report

(3 results)
  • 1998 Annual Research Report   Final Research Report Summary
  • 1997 Annual Research Report
  • Research Products

    (20 results)

All Other

All Publications (20 results)

  • [Publications] Toshihiro IHARA: "Gene Sensor Using Ferrocenyl Oligonucleotide" J. Chem. Soc, Chem. Commn.17. 1609-1610 (1997)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] Yoshiki KATAYAMA: "Cyclic AMP Detection by Electrode Modified with 17 mer Oligopeptide" Chem. Lett.1997. 883-884 (1997)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] Daisuke Umeno: "Single-stranded DNA-poly(N-isopropylacrylamide) Comjugate for Affinity Precipitation Separation of Oligonucleotides" J. Chem. Soc. Chem. Commun.1443-1444 (1998)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] Koji NAKANO: "DNA-Modified Electrode. Molecular Recognition and Electrochemical Response" ACS Symp. Ser.703. 34-45 (1998)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] Mizuo MAEDA: "Molecular and Ionic Recognition with Imprinted Polymers : A Brief Overview" ACS Symp. Ser.703. 1-8 (1998)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] Yoshiki KATAYAMA: "Bioaffinity Sensor to Anti-DNA Antibodies Using DNA Modified Au Electrode" Chem. Lett.1998. 1181-1182 (1998)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] Toshihiro Ihara, Masamichi Nakayama, Mitsuru Murata, Koji Nakano, Mizuo Maeda: "Gene Sensor Using Ferrocenyl Oligonucleotide" J.Chem.Soc.Chem.Commun.1609-1610 (1997)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] Yoshiki Katayama, Yuya Ohuchi, Mizuo Maeda, Hideyoshi Higashi, Yoshihisa Kudo: "Cylic AMP Detectiopn by Electrode Modified with 17mer Oligopeptide" Chem.Lett.883-884 (1998)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] Daisuke Umeno, Takeshi Mori, Mizuo Maeda: "Single-stranded DNA-poly (N-isopropylacrylamide) Comjugate for Affinity Precipitation Separation of Oligonucleotides" J.Chem.Soc.Chem.Commun.1443-1444 (1998)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] Koji Nakano, Shinji Uchida, Yoshiharu Mitsuhashi, Yuji Fujita, Hiroaki Taira, Mizuo Maeda: "DNA-Modified Electrode. Molecular Recognition and Electrochemical Response" ACS Symp.Ser.690. 34-45 (1998)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] Mizuo Maeda, Richard A.Bartsch: "Molecular and Ionic Recognition with Imprinted Polymers : A Brief Overview" ACS Symp.Ser.703. 1-8 (1998)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] Yoshiki Katayama, Masamichi Nakayama, Hiroshi Irie, Mizuo Maeda: "Bioaffinity Sensor to Anti-DNA Antibodies DNA Modified Au Electrode" Chem.Lett.1181-1182 (1998)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] Daisuke UMENO: "Single-stranded DNA-poly (N-isopropylaceylamide)conjugate for affinity precipitation separation of oligonucleotides" J.Chem.SOC,Chem.Commun.1998. 1433-1444 (1998)

    • Related Report
      1998 Annual Research Report
  • [Publications] Koji NAKANO: "DNA-Modified Electrode.Molecular Recognition and Electrochemical Response" American Chemical Society,ACS Symp.Ser.690. 34-45 (1998)

    • Related Report
      1998 Annual Research Report
  • [Publications] Mizuno Maeda: "Molecular and Ionic Recognition with Imprinted Polymers:A Brief Overview" American Chemical Society,ACS Symp.Ser.703. 1-8 (1998)

    • Related Report
      1998 Annual Research Report
  • [Publications] Yoshiki KATAYAMA: "Bioaffinity Sensor to Anti-DNA Antibodies Using DNA Modified Au Electrode" Chem.Lett.1998. 1181-1182 (1998)

    • Related Report
      1998 Annual Research Report
  • [Publications] Yoshiki KATAYAMA: "Cyclic AMP Detection by Electrode Modified with 17mer Oligopeptide" Chemistry Letters. 1997. 883-884 (1997)

    • Related Report
      1997 Annual Research Report
  • [Publications] Toshihiro IHARA: "Gene sensor using ferrocenyl oligonucleotide" J.Chem.Soc.,Chem.Commun.1997・17. 1609-1610 (1997)

    • Related Report
      1997 Annual Research Report
  • [Publications] 中山正道: "遺伝子センサ" 海外高分子研究. 43. 129-130 (1997)

    • Related Report
      1997 Annual Research Report
  • [Publications] 前田瑞夫: "DNAバイオセンサ" 化学と工業. 50・7. 992-994 (1997)

    • Related Report
      1997 Annual Research Report

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Published: 1997-04-01   Modified: 2016-04-21  

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