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DNA型および血液型の超高感度検出法の開発:とくに微量体液斑からの型判定

Research Project

Project/Area Number 16689015
Research Category

Grant-in-Aid for Young Scientists (A)

Allocation TypeSingle-year Grants
Research Field Legal medicine
Research InstitutionShimane University

Principal Investigator

竹下 治男  島根大学, 医学部, 教授 (90292599)

Project Period (FY) 2004 – 2006
Project Status Completed (Fiscal Year 2006)
Budget Amount *help
¥27,820,000 (Direct Cost: ¥21,400,000、Indirect Cost: ¥6,420,000)
Fiscal Year 2006: ¥8,060,000 (Direct Cost: ¥6,200,000、Indirect Cost: ¥1,860,000)
Fiscal Year 2005: ¥9,100,000 (Direct Cost: ¥7,000,000、Indirect Cost: ¥2,100,000)
Fiscal Year 2004: ¥10,660,000 (Direct Cost: ¥8,200,000、Indirect Cost: ¥2,460,000)
KeywordsDNA型 / 血液型 / 超高感度検出法 / 微量体液斑 / 型判定 / DNaseI / DNASE1多型 / HumDN1多型 / DNase I / PCR / 精製 / DAFO / 尿 / 尿斑
Research Abstract

最終年度にあたり、DNA型および血液型の超高感度検出法の開発、とくに微量体液斑について、個人識別能が高いうえ、経時変化に抵抗性を示す、デオキシリボ核酸分解酵素(DNaseI)型における実用化の検討やデータベース作成を行った。DNaseI多型において、既存のDNaseI(DNASE1)多型に加えて、最近、ヒトDNaseI遺伝子のイントロン4内に存在している反復塩基配列が新規なVNTR多型であることが解明され、HumDN1多型と命名された。しかし,ヒトDNaseIの多型頻度分布調査はこれまでのところ,日本人およびドイツ人の2集団に限られている.そこで本研究では,島根県在住の日本人、韓国人およびモンゴル人集団の微量血液、トルコ人およびアフリカオバンボス人集団の微量血痕由来DNAから,等電点電気泳動に基づく蛋白多型検出ではなく,新たな同時PCR法に基づくDNA多型同時検出法開発によりDNASE1およびHumDN1多型の頻度分布調査を行った。両多型とも、トルコ人集団の出現頻度はドイツ人集団に類似しており,DNASE1*2、HumDN1*4およびHumDN1*5の出現頻度は他集団に比べ有意に高かった.アフリカオバンボス人集団では、HumDN1*1およびHumDN1*3の出現頻度が他集団に比して有意に高かった。アジア人3集団においては、両多型とも類似した集団データを示した。DNaseI遺伝子に存在するDNASE1多型とHumDN1多型との間の相関について解析したところ、両多型の間に連鎖不平衡が認められた。いずれの集団においても、HumDN1*3はDNASE1*1と、HumDN1*4およびHumDN11*5はDNASE1*2と有意に相関していた。以上から、今回構築し得た微量体液斑からの両DNaseI多型検出法は、DNA型および血液型の超高感度検出法開発の一助になり得るものと考えられた。

Report

(3 results)
  • 2006 Annual Research Report
  • 2005 Annual Research Report
  • 2004 Annual Research Report
  • Research Products

    (28 results)

All 2007 2006 2005 2004

All Journal Article (28 results)

  • [Journal Article] Specific determination of linear alkylbenzenesulfonates (LAS) in commercial detergents and whole blood by high-performance liquid chromatography with solid phase extraction.2007

    • Author(s)
      Xue Y, Hieda Y, Fujihara J, Takayama K, Takeshita H
    • Journal Title

      Journal of Analytical Toxicology 31・1

      Pages: 37-43

    • Related Report
      2006 Annual Research Report
  • [Journal Article] Variation of interleukin 8-251 A>T polymorphism in worldwide populations and intra-ethnic differences in Japanese populations2007

    • Author(s)
      Fujihara J, Shiwaku K, Yasuda T, Yuasa I, Nishimukai H, Iida R, Takeshita H
    • Journal Title

      Clinica Chimica Acta 377・1-2

      Pages: 79-82

    • Related Report
      2006 Annual Research Report
  • [Journal Article] Two deoxyribonuclease I gene polymorphisms and correlation between genotype and its activity in Japanese population2007

    • Author(s)
      Fujihara J, Takatsuka H, Kataoka K, Xue Y, Takeshita H
    • Journal Title

      Legal Medicine (in press)

    • NAID

      10024172712

    • Related Report
      2006 Annual Research Report
  • [Journal Article] CYP1A2 polymorphism (C>A at position-163) in Ovambos, Koreans and Mongolians.2007

    • Author(s)
      Fujihara J, Shiwaku K, Xue Y, Kataoka K, Hieda Y, Takeshita H
    • Journal Title

      Cell Biochemistry and Function (in press)

    • Related Report
      2006 Annual Research Report
  • [Journal Article] Extremely high prevalence of DNASE1^*1 allele in African populations.2007

    • Author(s)
      Takeshita H, Fujihara J, Soejima M, Koda Y, Nakajima T
    • Journal Title

      Cell Biochemistry and Function (in press)

    • Related Report
      2006 Annual Research Report
  • [Journal Article] Allele frequencies for nine STR loci in Ovambo population using AmpFISTR^[○!R] Profiler Kit.2007

    • Author(s)
      Fujihara J, Muro T, Nakamura H, Imamura S, Takeshita H
    • Journal Title

      Forensic Science of International (in press)

    • Related Report
      2006 Annual Research Report
  • [Journal Article] Frequency of two human glutathione-S-transferase Ω1 polymorphisms (E155 deletion and E208K) in Ovambo and Japanese populations using the PCR-based genotyping method2007

    • Author(s)
      Fujihara J, Kunito T, Agusa T, Iwata H, Takeshita H
    • Journal Title

      Clinical Chemistry Laboratory Medicine 45・5(in press)

    • Related Report
      2006 Annual Research Report
  • [Journal Article] Actin-inhibition and folding of vertebrate deoxyribonuclease I are affected by mutations at residues 67 and 114.2006

    • Author(s)
      Fujihara J, Takeshita H et al.
    • Journal Title

      Comparative Biochemistry and Physiology Part B 143・1

      Pages: 70-75

    • Related Report
      2006 Annual Research Report 2005 Annual Research Report
  • [Journal Article] One-step purification of mammalian Deoxyribonucleases I, and differences among pancreas, parotid and pancreas-parotid (mixed) types based on species-and organ-specific N-linked glycosylation.2006

    • Author(s)
      Fujihara J, Takeshita H et al.
    • Journal Title

      Biochemistry (Moscow) 71・Suppl

    • Related Report
      2006 Annual Research Report
  • [Journal Article] Single-step purification by lectin affinity and deglycosylation analysis of recombinant human and porcine deoxyribonuclease I expressed in COS-7 cells.2006

    • Author(s)
      Fujihara J, Takeshita H et al.
    • Journal Title

      Biotechnology Letters 28・4

      Pages: 215-221

    • Related Report
      2006 Annual Research Report
  • [Journal Article] CYP2A6 polymorphism reveals differences in Japan and the existence of a specific variant in Ovambo and Turk populations.2006

    • Author(s)
      Takeshita H et al.
    • Journal Title

      Human Biology 78・2

      Pages: 235-242

    • Related Report
      2006 Annual Research Report
  • [Journal Article] Analysis of genetic polymorphism of deoxyribonuclease I in Japanese from Shimane prefecture using genotyping method.2006

    • Author(s)
      Takatsuka H, Fujihara J, Xue Y, Imamura S, Takayama K, Kataoka K, Takeshita H
    • Journal Title

      Shimane Journal of Medical Science 23

      Pages: 1-6

    • NAID

      120005585551

    • Related Report
      2006 Annual Research Report
  • [Journal Article] Fequency of a single nucleotide (A2317G) and 56-bp variable number of tandem repeat polymorphisms within the deoxyribonuclease I gene in five ethnic populations2006

    • Author(s)
      Fujihara J, Yasuda T, Shiwaku K, Takeshita H
    • Journal Title

      Clinical Chemistry Laboratory Medicine 44・10

      Pages: 1188-1191

    • Related Report
      2006 Annual Research Report
  • [Journal Article] One-step purification of mammalian Deoxyribonucleases I, and differences among pancreas, parotid and pancreas-parotid (mixed) types based on species-and organ-specific N-linked glycosylation.2006

    • Author(s)
      Fujihara J, Takeshita H et al.
    • Journal Title

      Biochemistry (Moscow) 71

    • Related Report
      2005 Annual Research Report
  • [Journal Article] Single-step purification by lectin affinity and deglycosylation analysis of recombinant human and porcine deoxyribonuclease I expressed in COS-7 cells.2006

    • Author(s)
      Fujihara J, Takeshita H et al.
    • Journal Title

      Biotechnology Letters (in press)

    • Related Report
      2005 Annual Research Report
  • [Journal Article] CYP2A6^*1A, ^*1B, ^*1F, ^*1G ^*4A and ^*4D polymorphism reveals differences among small areas of Japan and the existence of a specific variant in Ovambo and Turk populations.2006

    • Author(s)
      Takeshita H et al.
    • Journal Title

      Human Biology (in press)

    • Related Report
      2005 Annual Research Report
  • [Journal Article] CYP1A2polymorphism (C>A at position-163) in Ovambos, Koreans and Mongolians.2006

    • Author(s)
      Fujihara J, Takeshita H et al.
    • Journal Title

      Cell Biochemistry and Function (in press)

    • Related Report
      2005 Annual Research Report
  • [Journal Article] A fatal case of pure ethanol ingestion.2005

    • Author(s)
      Hieda Y, Takeshita H et al.
    • Journal Title

      Forensic Science International 149・2/3

      Pages: 243-247

    • Related Report
      2005 Annual Research Report
  • [Journal Article] Skin analysis to determine causative agent in dermal exposure to petroleum products.2005

    • Author(s)
      Hieda Y.Takeshita H et al.
    • Journal Title

      Journal of Chromatography B 823・1

      Pages: 53-59

    • NAID

      10016979736

    • Related Report
      2005 Annual Research Report
  • [Journal Article] Analysis of genetic polymorphism of deoxyribonuclease I (DNase I) in Ovambo and Turk populations using genotyping method.2005

    • Author(s)
      Fujihara J, Takeshita H et al.
    • Journal Title

      Biochemical Genetics 43・11/12

      Pages: 631-638

    • Related Report
      2005 Annual Research Report
  • [Journal Article] GC-MS分析による皮膚分析の有用性と臨床および法医実務への応用2005

    • Author(s)
      稗田洋子, 竹下治男ら
    • Journal Title

      日本医用マススペクトル学会サーキュラー 65

      Pages: 2-7

    • Related Report
      2005 Annual Research Report
  • [Journal Article] A fatal case of pure ethanol ingestion.2005

    • Author(s)
      Hieda Y et al.
    • Journal Title

      Forensic Sci Int (in press)

    • Related Report
      2004 Annual Research Report
  • [Journal Article] Deoxyribonucleases I typing from tears.2004

    • Author(s)
      Fujihara J, Takeshita H et al.
    • Journal Title

      Shimane Journal of Medical Science 22・2

      Pages: 45-49

    • NAID

      120005582408

    • Related Report
      2005 Annual Research Report
  • [Journal Article] Rapid quantification of DNase I activity in one-microliter serum samples.2004

    • Author(s)
      Takeshita H et al.
    • Journal Title

      Clin Chem 50

      Pages: 446-448

    • Related Report
      2004 Annual Research Report
  • [Journal Article] Distribution and disposition of benzalkonium chloride following various routes of administration in rats.2004

    • Author(s)
      Xue Y et al.
    • Journal Title

      Toxicol Lett 148(1/2)

      Pages: 113-123

    • Related Report
      2004 Annual Research Report
  • [Journal Article] A single amino acid substitution can shift the optimum pH of DNase I for enzyme activity : biochemical and molecular analysis of the piscine DNase I family.2004

    • Author(s)
      Takeshita H et al.
    • Journal Title

      Biochim Biophys Acta 1672(3)

      Pages: 174-183

    • Related Report
      2004 Annual Research Report
  • [Journal Article] A novel 56-bp variable tandem repeat polymorphism in the human deoxyribonuclease I gene and its population data.2004

    • Author(s)
      Yasuda T et al.
    • Journal Title

      Legal Med 6(4)

      Pages: 242-245

    • Related Report
      2004 Annual Research Report
  • [Journal Article] Molecular evolution of shark and other vertebrate DNases I.2004

    • Author(s)
      Yasuda T et al.
    • Journal Title

      Eur J Biochem 271(22)

      Pages: 4428-4435

    • Related Report
      2004 Annual Research Report

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

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