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Gene Detection by Light-to-electric Converting Intercalator and Its Application to DNA

Research Project

Project/Area Number 12305054
Research Category

Grant-in-Aid for Scientific Research (A)

Allocation TypeSingle-year Grants
Section一般
Research Field 工業分析化学
Research InstitutionKyushu University

Principal Investigator

TAKAGI Makoto  Faculty of Engineering, Prof., 大学院・工学研究院, 教授 (90037739)

Co-Investigator(Kenkyū-buntansha) NOJIMA Takahiko  Faculty of Engineering Research Associate, 大学院・工学研究院, 助手 (00291930)
NAKAMURA Shigeo  Miyazaki Univ., Faculty of Engineering, Assoc. Prof., 工学部, 助教授 (00264078)
TAKENAKA Shigeori  Faculty of Engineering, Assoc. Prof., 大学院・工学研究院, 助教授 (60188208)
Project Period (FY) 2000 – 2002
Project Status Completed (Fiscal Year 2002)
Budget Amount *help
¥23,960,000 (Direct Cost: ¥20,600,000、Indirect Cost: ¥3,360,000)
Fiscal Year 2002: ¥4,940,000 (Direct Cost: ¥3,800,000、Indirect Cost: ¥1,140,000)
Fiscal Year 2001: ¥9,620,000 (Direct Cost: ¥7,400,000、Indirect Cost: ¥2,220,000)
Fiscal Year 2000: ¥9,400,000 (Direct Cost: ¥9,400,000)
Keywordsphotocurrent / gene detection / DNA chip / intercalator / ruthenium complex / threading intercalation
Research Abstract

In this research we developed a novel detection system of DNA duplex using a light-to-electric conversion. The principle of the detection of DNA duplex formation is as follows. 1) The gold electrode modified by single stranded DNA was prepared. 2) When double stranded DNA is formed, ruthenium complex will intercalate and will be concentrated on the electrode surface. 3) Then the visible light is irradiated to the electrode, photocurrent caused by excitation of ruthenium complex will be observed.
We used a polypyridyl ruthenium complex carrying DNA intercalating ligand for DNA detection. Ru(bpy)_3 was used as a control compound. Ru(bpy)_2dppz was used as a ruthenium complex carrying a DNA intercalating part. Furthermore, we synthesized NDI-Ru_2 as a threading intercalator carrying ruthenium complex. This type of compound has a smaller dissociation rate from DNA duplex, because it has threading intercalating part connected to two bulky groups
A DNA-immobilized gold electrode was dipped into the solution containing the light-to-electric converting intercalator (Ru(bpy)_3, Ru(bpy)_2dppz, NDI-Ru_2). When the visible light was irradiated to the surface of the electrode, continuous photocurrent occurred in the presence of tertiary amine as an electron donor. The amount of photocurrent increased by 36% in the case of Ru(bpy)_2dppz, 86% in the case of NDI-Ru_2 after the formation of double-stranded DNA. This result suggested that the metallointercalator was concentrated on the surface of gold electrode by its affinity to DNA duplex. In the case of NDI-Ru_2, we consider that electron was smoothly transferred from ruthenium to gold electrode through naphthalenediimide because NDI acts as an electron acceptor. In conclusion, we could discriminate between double stranded and single stranded DNA from the magnitude of the photocurrent using a ruthenium intercalator.

Report

(4 results)
  • 2002 Annual Research Report   Final Research Report Summary
  • 2001 Annual Research Report
  • 2000 Annual Research Report
  • Research Products

    (27 results)

All Other

All Publications (27 results)

  • [Publications] B.Juskowiak, et al.: "Fluorescence energy transfer study of interstrand DNA cross-linking caused by rigid bisintercalator"Supramolecular Chemistry. 14. 477-485 (2002)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] S.Nakamura, et al.: "Light-to-electric conversion as a new method for DNA duplex detection"Analytical Sciences. 17. 431-432 (2001)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] B.Juskowiak, et al.: "Isomerization of DNA-bound distilbazolium ligand induced by electron transfer photoexcited tris(1,10-phenanthroline)Ru(II)"Photochemistry and Photobiology. 74. 391-400 (2001)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] A.Kumasaki, et al.: "A cyclic sugar unit interacts more strongly with DNA than do open-chain polyol coun erparts in sugar-linked synthetic DNA intercalators"Chemistry Letters. 2001. 370-371 (2001)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] B.Mathew, et al.: "Lipophilic peptide as novel sensory material for chiral amine recognition"Chemistry Letters. 2000. 1029-1030 (2000)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] T.Kuboyama, et al.: "Lipophilic peptide as novel building material of molecular sensor"Chemistiy Letters. 2000. 552-553 (2000)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] B.Juskowiak, et al.: "Fluorescence energy transfer study of interstrand DNA cross-linking caused by rigid bisintercalator"Supramolecular Chemistry. 14. 477-485 (2002)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] S.Nakamura, et al.: "Light-to-electric conversion as a new method for DNA duplex detection"Analytical Sciences. 17. 431-432 (2001)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] B.Juskowiak, et al.: "Isomerization of DNA-bound distilbazolium ligand induced by electron transfer photoexcited tris(1, 10-phenanthroline)Ru(II)"Photochemistry and Photobiology. 74. 391-400 (2001)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] A.Kumasaki, et al.: "A cyclic sugar unit interacts more strongly with DNA than do open-chain polyol counterparts in sugar-linked synthetic DNA intercalators"Chemistry Letters. 370-371 (2001)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] B.Mathew et al.: "Lipophilic peptide as novel sensory material for chiral amine recognition"Chemistry Letters. 1026-1027 (2000)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] T.Kuboyama et al.: "Lipophilic peptide as novel building material of molecular sensor"Chemistry Letters. 552-553 (2000)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] K.Yamashita, M.Takagi, H.Kondo, S.Takenaka: "Electrochemical detection of nucleic base mismatches with ferrocenyl naphthalene diimide"Analytical Biochemistry. Vol. 306. 188-196 (2002)

    • Related Report
      2002 Annual Research Report
  • [Publications] H.Miyahara, K.Yamashita, M.Kanai, K.Uchida, M.Takagi, H.Kondo, S.Takenaka: "Electrochemical analysis of single nucleotide polymorphisms of p53 gene"Talanta. Vol. 56. 829-835 (2002)

    • Related Report
      2002 Annual Research Report
  • [Publications] S.Sato, T.Nojima, M.Takagi, H.Kondo, S.Takenaka: "Synthesais of adamantly naphthalene diimide and its interaction with double stranded DNA"Nucleic Acids Research Supplement. 2. 213-214 (2002)

    • Related Report
      2002 Annual Research Report
  • [Publications] S.Fujii, N.Okimoto, T.Ebisuzaki, T.Nojima, M.Takagi: "Molecular dynamics investigation of the double stranded oligonucleotide d(AT)6d(AT)6"Nucleic Acids Research Supplement. 2. 175-176 (2002)

    • Related Report
      2002 Annual Research Report
  • [Publications] H.Torigoe, S.Sato, K.Yamashita, S.Okiba, T.Imanishi, S.Takenaka: "Binding of threading intercalator to nucleic acids : Thermodynamic analysis"Nucleic Acids Research Supplement. 2. 55-56 (2002)

    • Related Report
      2002 Annual Research Report
  • [Publications] S.Takenaka: "Small Molecule DNA and RNA Binders : From Synthesis to Nucleic Acid"Wiley-VCH. 23 (2003)

    • Related Report
      2002 Annual Research Report
  • [Publications] S.Nakamura et al.: "Light-to-Electric Conversion as a New Method for DNA Duplex Detection"Anal. Sci.. (in press).

    • Related Report
      2001 Annual Research Report
  • [Publications] H.Miyahara et al.: "Electrochemical array (ECA) as an integrated multi-electrode DNA sensor"Trans. IEE of Japan. 121-E・4. 187-191 (2001)

    • Related Report
      2001 Annual Research Report
  • [Publications] S.Takenaka: "Highly sensitive probe for gene analysis by electrochemical approach"Bull. Chem. Soc. Jpn. 74・2. 217-224 (2001)

    • Related Report
      2001 Annual Research Report
  • [Publications] K.Yamashita et al.: "Visualization of DNA micro arrays by scanning electrochemical microscopy (SECM)"Analyst. 126. 1201-1202 (2001)

    • Related Report
      2001 Annual Research Report
  • [Publications] M, Takagi: "Threading intercalation to double-stranded DNA and the application to DNA sensing. Electrochemical array technique"Pure Appl. Chem.. 73・10. 1573-1577 (2001)

    • Related Report
      2001 Annual Research Report
  • [Publications] S.Takenaka et al.: "Base mutation analysis by a ferrocenyl naphthalene diimide drivative"Nucleotides & Nucleic Acids. 20・4・7. 1429-1432 (2001)

    • Related Report
      2001 Annual Research Report
  • [Publications] 竹中繁織: "分析化学便覧 h.DNAおよびRNA"丸善. 7 (2001)

    • Related Report
      2001 Annual Research Report
  • [Publications] S.Takenaka et al.: "DNA Sensing on a DNA Probe-Modefied Electrode Using Ferrocenylnaphthalene Diimide as the Electrochemically Active Ligand"Analytical Chemistry. 72(6). 1334-1341 (2000)

    • Related Report
      2000 Annual Research Report
  • [Publications] K.Yamashita et al.: "Electrochemical Detection of Base Pair mutation"Chemistry Letters. (9). 1038-1039 (2000)

    • Related Report
      2000 Annual Research Report

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Published: 2000-04-01   Modified: 2016-04-21  

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