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STUDY FOR MECHANISM OF TRIPLEX USING SINGLE MOLECULES DETECTION

Research Project

Project/Area Number 07808072
Research Category

Grant-in-Aid for Scientific Research (C)

Allocation TypeSingle-year Grants
Section一般
Research Field Biophysics
Research InstitutionHOKKAIDO UNIVERSITY

Principal Investigator

KINJO Masataka  Hokkaido Univ.Res.Inst.Elect.Sci., Res.Assoc., 電子科学研究所, 助手 (70177971)

Co-Investigator(Kenkyū-buntansha) NISHIMURA Goro  Hokkaido Univ.Res.Inst.Elect.Sci., Res.Assoc., 電子科学研究所, 助手 (30218193)
Project Period (FY) 1995 – 1996
Project Status Completed (Fiscal Year 1996)
Budget Amount *help
¥2,100,000 (Direct Cost: ¥2,100,000)
Fiscal Year 1996: ¥500,000 (Direct Cost: ¥500,000)
Fiscal Year 1995: ¥1,600,000 (Direct Cost: ¥1,600,000)
KeywordsDNA / TRIPLEX / FLUORESCENCE CORRELATION SPECTROSCOPY / FLUORESCENCE / AUTOCORRELATION / RHODAMIN / SINGLE MOLECULE / HYBRIDIZATION / 分子量
Research Abstract

PURPOSE :
The formation process of triple helix DNA in an aqueous solution was analyzed using Fluorescence Correlation Spectroscopy in tiny volume at the single molecule detection. The triple helix DNA was made with fluorescence labeled oligo T15 as a probe DNA strand and homo purin : homo pyrimidine DNA as a target strand. Changing the translational diffusion time of probe DNA was used as indicator for the rate of producing of triplex.
We confirmed the FCS could be used as determination method for triplex in the first budget year, and we have applied the FCS method to analysis the formation of triplex in solution at the second budget year.
METHODS :
Various length of fluorescence labeled DNA (50bp to 7000bp) was synthesized using PCR method in the presence of fluorescence tagged dUTP (Rho-dUTP). The translational diffusion time of these DNA were measured with FCS and had been used as standard. In order to compare with the formation process of triplex, we measured the associate process of T15 to complementary strand DNA.
CONCLUSION :
The translational diffusion time of double strand DNA agreed with the theoretical values for a rod-shaped model. We could analyze the translational diffusion time of triplex DNA using same method with two components model since triplex has more rigid structure than double strand DNA.The results show the formation rate of triplex was slower than that of double helix. Also, about 80% of probe DNA bind to target double strand DNA while all of added probe DNA strand bind to target DNA in the case of double strand DNA.
The experiment with mismatch sequences in the target and/or the probe DNA will be analyzed in future.

Report

(3 results)
  • 1996 Annual Research Report   Final Research Report Summary
  • 1995 Annual Research Report
  • Research Products

    (42 results)

All Other

All Publications (42 results)

  • [Publications] Kinjo M.: "Ultrasensitive hybridization analysis using fluorescence correlation spectroscopy." Nucleic Acid Res.23. 1795-1799 (1995)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Katayama K.: "Absorbance measurement in turbid media by photon correlation method" Applied Optics. 34. 7419-7427 (1995)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] M.Wakita: J. Biochem. (Tokyo).118. 1151-1160 (1995)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] M.Oda: "Determination of absolute concentration of oxy-and deoxyhemoglobin in rat head by Time-Resolved Beer-Lambert Law." SPIE. 2389. 770-778 (1995)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] 金城政孝: "拡散測定によるDNA構造解析-レーザー自己相関蛍光法の開発" 日本バイオレオロジー学会誌. 第9巻. 74-83 (1995)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] 金城政孝: "光を使った分子診断,相関分光法による単一分子の検出" 化学. 第50巻. 556-559 (1995)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Nishimura G.: "Diffusing-Wave Absorption Spectroscopy in the Homegeneous Turbid Media," Opt. Commun.128. 99-107 (1996)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Thyberg P.: "Global analysis of molecular interactions : Binding of a primer to DNA" Prog. Biophys. Molec. Biol.65. 81-81 (1996)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Nisimura G.: "Analysis of FCS data of rhodamine labled dsDNA under cleaving reaction." Prog. Biophys. Molec. Biol.65. 197-197 (1996)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Kinjo M.: "Analysis fo the munber of DNA fragment in solution using fluorescence correlation spectroscopy." Prog. Biophys. Molec. Biol.65. 197-197 (1996)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] 金城政孝: "自己相関蛍光法による制限酵素断片数の評価。" 電子科学研究. 第3巻. 75-77 (1995)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] 金城政孝: "遺伝子の動きを見る。" 電子科学研究. 第4巻. 96-98 (1996)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Kinjo M.& Rigler R: "Ultrasensitive hybridization analysis using fluorescence correlation spectroscopy." Nucleic Acid Res. 23 (10). 1795-1799 (1995)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Katayama K., Nishimura G., Kinjo M.& Tamura M: "Absorbance measurement in turbid media by photon correlation method" Applied Optics. 34. 7419-7427 (1995)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] M.Wakita, G.Nishimura and M.Tamura: "Some Characteristics of the Fluorescence Lifetime of Reduced Pyridine Nucleotides in Isolated Mitochondria, Isolated Hepatocyte, and Perfused Liver of Rat In situ." J.Biochem. (Tokyo). 118. 1151-1160 (1995)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] M.Oda, Y.Yamashita, G.Nishimura and M.Tamura: "Determination of absolute concentration of oxy-and deoxyhemoglobin in rat head by Time-Resolved Beer-Lambert Law." SPIE. 2389. 770-778 (1995)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Kinjo M.& Koyama T.: "Analysis of DNA structure by measurement of diffusion rate -Development of fluorescence correlation spectroscopy with continuous wave laser-." J.Jpn Soc.Biorheology. 9 (2). 74-83 (1995)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Nishimura G., Katayama K., Kinjo M.& Tamura M.: "Diffusing-Wave Absorption Spectroscopy in the Homegeneous Turbid Media." Opt.Commun. 128. 99-107 (1996)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Jin T., Kinjo M., Koyama T., Kobayashi Y.& Hitata H.: "Selective Na+ transport through phosopholipid bilayr membrne by a synthetic Calix [4] arene carrier." Langmuir. 12. 2684-2689 (1996)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Thyberg P., Kinjo M.& Rigler R.: "Global analysis of molecular interactions : Binding of a primer to DNA" Prog.Biophys.Molec.Biol. 65. 81 (1996)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Nisimura G., Rigler R.& Kinjo M.: "Analysis of FCS data of rhodamine labled dsDNA under cleaving reaction." Prog.Biophys.Molec.Biol. 65. 197 (1996)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Kinjo M.& Nishimura G.: "Analysis fo the munber of DNA fragment in solution using fluorescence correlation spectroscopy." Prog.Biophys.Molec.Biol. 65. 197 (1996)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Kinjo Masataka: "Jikosoukann bunnkouhou ni yoru seigenn kouso dannpenn suu no hyouka." Dennshi Kagaku Kennkyuu. 3. 75-77 (1995)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Kinjo Masataka: "Idennshi no ugoki wo miru." Dennshi Kagaku Kennkyuu. 4. 96-98 (1996)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Kinjo M.: "Ultrasensitive hybridization analysis using fluorescence correlation spectroscopy." Nucleic Acid Res.23(10),. 1795-1799 (1995)

    • Related Report
      1996 Annual Research Report
  • [Publications] Katayama K.: "Absorbance measurement in turbid media by photon correlation method" Applied Optics. 34,. 7419-7427 (1995)

    • Related Report
      1996 Annual Research Report
  • [Publications] M. Wakita: "Some Characteristics of the Fluorescence Lifetime of Reduced Pyridine Nucleotides inIsolated Mitochondria, Isolated Hepatocyte, and Perfused Liver of Rat In situ," J. Biochem.(Tokyo). 118. 1151-1160 (1995)

    • Related Report
      1996 Annual Research Report
  • [Publications] M. Oda,: "Determination of absolute concentration of oxy-and deoxyhemoglobin in rat head by Time-Resolved Beer-Lambert Law," SPIE. 2389. 770-778 (1995)

    • Related Report
      1996 Annual Research Report
  • [Publications] 金城政孝: "拡散測定によるDNA構造解析-レーザー自己相関蛍光法の開発-" 日本バイオレオロジー学会誌. 第9巻. 74-83 (1995)

    • Related Report
      1996 Annual Research Report
  • [Publications] 金城政孝: "光を使った分子診断、相関分光法による単一分子の検出" 化学. 第50巻,. 556-559 (1995)

    • Related Report
      1996 Annual Research Report
  • [Publications] Nishimura G.: "Diffusing-Wave Absorption Spectroscopy in the Homegeneous Turbid Media," Opt. Commun.128,. 99-107 (1996)

    • Related Report
      1996 Annual Research Report
  • [Publications] Thyberg P.: "Global analysis of molecular interactions : Binding of a primer to DNA" Prog. Biophys. Molec. Biol.65. 81-81 (1996)

    • Related Report
      1996 Annual Research Report
  • [Publications] Nishimura G.: "Analysis of FCS data of rhodamine labled dsDNA under cleaving reaction." Prog. Biophys. Molec. Biol.65. 197-197 (1996)

    • Related Report
      1996 Annual Research Report
  • [Publications] Kinjo M.: "Analysis fo the munber of DNA fragment in solution using fluorescence correlation spectroscopy." Prog. Biophys. Molec. Biol.65. 197-197 (1996)

    • Related Report
      1996 Annual Research Report
  • [Publications] 金城政孝: "自己相関蛍光法による制限酵素断片数の評価。" 電子科学研究. 第3巻. 75-77 (1995)

    • Related Report
      1996 Annual Research Report
  • [Publications] 金城政孝: "遺伝子の動きを見る。" 電子科学研究. 第4巻. 96-98 (1996)

    • Related Report
      1996 Annual Research Report
  • [Publications] M.Kinjo and R.Rigler: "Ultrasensitive hybridization analysis using fluorescena corvelation Spectroscopy" Nueleic Acid Research. 23. 1795-1799 (1995)

    • Related Report
      1995 Annual Research Report
  • [Publications] K.Katayama,G.Hishimura,M.Kinjo and M.tamura: "Absorption measurement in turbid by proton correlation method," Applied Opt.34. 7419-7427 (1995)

    • Related Report
      1995 Annual Research Report
  • [Publications] G.Nishimura,K.Katayama,M.Kinjo,and M.Tamura: "Diffusing-Wave absorption spoctroscopy in the homegeneous turbid media" Opt.Commun.(accepted for publication). (1996)

    • Related Report
      1995 Annual Research Report
  • [Publications] M.Oda,Y.Yamashita,G.Nishimura,and T.Tamura: "Determination of absolute concentration of oxy-ovol deoxy hemoglobin in rat head by time-resolvod Beer-lambort low" S.P.I.E.2389. 770-778 (1995)

    • Related Report
      1995 Annual Research Report
  • [Publications] 金城政孝・小宮富康: "拡散測定によるDNA構造解析-レーザー自己相関蛍光測定法の解発" 日本バイオレオロジー学会誌. 9. 74-83 (1995)

    • Related Report
      1995 Annual Research Report
  • [Publications] 金城政孝: "光を使った分子診断、相関分光法による単一分子の検出" 化学. 50. 556-559 (1995)

    • Related Report
      1995 Annual Research Report

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Published: 1995-04-01   Modified: 2016-04-21  

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