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Basic Studies on DNA Polymorphism for Application to Foresic Medicine

Research Project

Project/Area Number 01480212
Research Category

Grant-in-Aid for General Scientific Research (B)

Allocation TypeSingle-year Grants
Research Field Legal medicine
Research InstitutionUniversity of Tsukuba

Principal Investigator

MISAWA Shogo  Univ. of Tsukuba, Inst. Community Med. Sci., Prof., 社会医学系, 教授 (50086534)

Co-Investigator(Kenkyū-buntansha) TANAKA Einosuke  Univ. of Tsukuba, Inst. Community Med. Sci., Assist. Prof., 社会医学系, 講師 (30138416)
HARADA Shoji  Univ. of Tsukuba, Inst. Community Med. Sci., Associate Prof., 社会医学系, 助教授 (60086618)
Project Period (FY) 1989 – 1990
Project Status Completed (Fiscal Year 1990)
Budget Amount *help
¥6,100,000 (Direct Cost: ¥6,100,000)
Fiscal Year 1990: ¥1,500,000 (Direct Cost: ¥1,500,000)
Fiscal Year 1989: ¥4,600,000 (Direct Cost: ¥4,600,000)
KeywordsDNA Polymorphism / DNA Fingerprint / Probe / RFLPs / Southern hybridization / Mitochondrial DNA / PCR / VNTR / RFLPs(制限酵素断片長多型) / サザンプロッテイング / PCR法 / 制限酵素 / サザンハイブリダイゼイション / Ha-ras遺伝子 / 個人性別 / 親子鑑定
Research Abstract

(1) Effects of Cytosine Methylation at Restriction Sites on Deoxyribonucleic Acid (DNA) Typing
The effects of endogenous 5-mrthylcytosines on deoxyribonucleic acid (DNA) fingerprints were studied. Analysis with methylation-sensitive restriction endonuclease Sau3AI and its methylation-insensitive isoschizomer MboI showed some differences in the patterns generated as a result of 5-methylcytosines at the recognition sites. Moreover a few bands of sperm DNA did not match those of blood DNA from the same individual, a phenomenon only observed in the digests of methylation-sensitive endonucleases. These findings indicate the unsuitability of methylation-sensitive restriction endonucleases for DNA fingerprinting and other forms of DNA typing, because of tissue-specific status of the methylation.
(2) Sex Identification by Analysis of DNA Extracted from Hard Tissues
Sex identification of unidentified two bleached skeletons by the method of DNA analysis in addition to the morphological examination … More was performed. Bones and teeth were taken out for the DNA analysis, and cracked to compounds by liquid nitrogen. DNA was extracted by the standard procedure. We used two methods for sex identification by DNA analysis ; one is Southern blot hybridization with Y-chromosome specific probe (PHY10 ; 3.4kb), and the other is PCR (polymerase chain reaction) amplifying with sex chromosome specific fragments (alphoid satellite family). These two methods enable us to identify the sex of the two cases presented here more correctly with sufficient evidence than depending only on morphological examination. With some technical difficulties, it is possible to put DNA analysis including Southern blot hybridization or PCR into practical determination of seing of hard tissues such as a piece of bone or a tooth, and old samples. Therefore, they seem to be effective methods applicable to forensic science.
(3) Probe walking : development of novel probes for DNA fingerprinting
The tanderm repeat of a 28-base-pair (bp) sequence downstream of the human c-Ha-ras-1 oncogene was studied as a probe for DNA fingerprinting. Multiple hypervar able patterns were observed by Southern hybridization at low stringency. The patterns were specific to individuals, we cloned the tandem repeat of a 33-bp sequence, which cross-hybridized with the 28-bp repeat. This 33-bp repeat detected another set of hypervariable restriction fragments by Southern hybridization at the same stringency. These results suggests that "prob walking" can be employed to develop novel probes that provide different DNA fingerprints. Less

Report

(3 results)
  • 1990 Annual Research Report   Final Research Report Summary
  • 1989 Annual Research Report
  • Research Products

    (36 results)

All Other

All Publications (36 results)

  • [Publications] Keiko Washio: "Effects of Cytosine Methylation at Restriction Sites on DeoxyribonucleicAcid(DNA)Typing" J. Forensic Sciences. 35. 1277-1283 (1990)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1990 Final Research Report Summary
  • [Publications] 本田 克也: "硬組織から抽出したDNAによる性別判定" 日本法医学雑誌. 44. 293-301 (1990)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1990 Final Research Report Summary
  • [Publications] 三木 敏行: "死亡した夫婦の実子の検査により父子及び母子鑑定例ー半同胞の鑑定例ー" Act. Crim. Japon.56. 263-274 (1990)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1990 Final Research Report Summary
  • [Publications] Shoji Harada: "Genetic Polymorphism of Aldehyde Dehydrogenase and its Physiological Significance to Alcohol Metabolism" Prog. Clin. Biol. Res.344. 289-294 (1990)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1990 Final Research Report Summary
  • [Publications] 原田 勝二: "アルコ-ル代謝酵素のDNAによる解析" アルコ-ル代謝と肝. 9. 23-26 (1990)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1990 Final Research Report Summary
  • [Publications] Shoji Harada: "Genetic Polymorphism of Aldehyde Dehydrogenase in Orientals as Genetic Marker of a High Risk Group of Alcoholism" Alcoholism and Family. 67-71 (1990)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1990 Final Research Report Summary
  • [Publications] 原田 勝二: "急性エタノ-ル中毒ー基礎編ー" 中毒研究. 3. 355-361 (1990)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1990 Final Research Report Summary
  • [Publications] 原田 勝二: "アルコ-ル耐性の個人差" 病態生理. 9. 948-955 (1990)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1990 Final Research Report Summary
  • [Publications] 原田 勝二: "エタノ-ルおよびエルデヒド代謝の人種的遺伝的要因" 医学のあゆみ. 154. 817-822 (1990)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1990 Final Research Report Summary
  • [Publications] 原田 勝二: "Glutathion SーTransferase 1 gene deletion およびApolipoprotein Bー100 gene 3'hypervariable regionのPCR法による解析とその応用" 2. 132-137 (1990)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1990 Final Research Report Summary
  • [Publications] Shoji Harada,: "Biological and Haematological Markers of" Pergamon Press Inc.,. 238-255 (1989)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1990 Final Research Report Summary
  • [Publications] Shoji Harada,: "Polymorphism of Aldehyde Dehydrogenase and its Application to Alcoholism" Electrophoresis. 10,. 309-318 (1989)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1990 Final Research Report Summary
  • [Publications] Surjit Singh, Shoji Harada,: "Inheritance for Mitochondrial Aldehyde Dehydrogenase ; Genotyping in Chinese, Japanese and South Korean Families Reveals Dominace of the Mutant Allele" Hum. Genet.83,. 119-121 (1989)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1990 Final Research Report Summary
  • [Publications] Keiko Washio, Shogo Misawa et al.,: "Individual Identification of Non-human Primates Using DNA Fingeprinting" Primates,. 30,. 217-221 (1989)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1990 Final Research Report Summary
  • [Publications] Keiko Washio, Shogo Misawa et al.,: "Probe Walking ; Development of Novel Probes for DNA Fingerprinting" Hum. Genet.,. 83,. 223-226 (1989)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1990 Final Research Report Summary
  • [Publications] Shinji Harihara, Shoji Harada, Shogo Misawa et al.,: "Family Study on Mitochondrial DNA Polymorphism" Jpn. J. Legal Med.,. 43,. 257-261 (1989)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1990 Final Research Report Summary
  • [Publications] Keiko Washio, Shogo Misawa et al.,: "Effects of Cytosine Methylation at Restriction Sites on Deoxyribonucleic Acid (DNA) Typing" J. Forensic Sciences,. 35,. 1277-1283 (1990)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1990 Final Research Report Summary
  • [Publications] Katsuya Honda, Shogo Misawa et al.,: "Sex Identification by Analysis of DNA Extracted from Hard Tissues" Jpn. J. Legal Med.,. 44,. 293-301 (1990)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1990 Final Research Report Summary
  • [Publications] Toshiyuki Miki, Shoji Harada et al.,: "A Case of Paternity and Maternity Where Putative Father and Putative Mother Were Deceased" Act. Crim. Japon.56,. 263-274 (1990)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1990 Final Research Report Summary
  • [Publications] Shoji Harada,: "Genetic Polymorphism of Aldehyde Dehydrogenase and its Physiological Significance to Alcohol Metabolism" Prog. Clin. Biol. Res.,. 344,. 289-294 (1990)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1990 Final Research Report Summary
  • [Publications] Shoji Harada,: "Genetic Polymorphism of Aldehyde Dehydrogenase in Orientals as Genetic Marker of a High Risk Group of Alcoholism" Alcoholism and Family,. 67-71 (1990)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1990 Final Research Report Summary
  • [Publications] Keiko Washio: "Effects of Cytosine Methylation at Restriction Sites on Deoxyribonucleic Acid (DNA) Typing" J.Forensic Sciences. 35. 1277-1283 (1990)

    • Related Report
      1990 Annual Research Report
  • [Publications] 本田 克也: "硬組織から抽出したDNAによる性別判定" 日本法医学雑誌. 44. 293-301 (1990)

    • Related Report
      1990 Annual Research Report
  • [Publications] 三木 敏行: "死亡した夫婦の実子の検査による父子及び母子鑑定例ー半同胞の鑑定例ー" Act.Crim.Japon.56. 263-274 (1990)

    • Related Report
      1990 Annual Research Report
  • [Publications] Shoji Harada: "Genetic Polymorphism of Aldehyde Dehydrogenase and its Physiological Significance to Alcohol Metabolism" Prog.Clin.Biol.Res.344. 289-294 (1990)

    • Related Report
      1990 Annual Research Report
  • [Publications] 原田 勝二: "アルコ-ル代謝酵素のDNAによる解析" アルコ-ル代謝と肝. 9. 23-26 (1990)

    • Related Report
      1990 Annual Research Report
  • [Publications] Shoji Harada: "Genetic Polymorphism of Aldehyde Dehydrogenase in Orientals as Genetic Marker of a High Risk Group of Alcoholism" Alcoholism and Family. 67-71 (1990)

    • Related Report
      1990 Annual Research Report
  • [Publications] 原田 勝二: "急性エタノ-ル中毒ー基礎編ー" 中毒研究. 3. 355-361 (1990)

    • Related Report
      1990 Annual Research Report
  • [Publications] 原田 勝二: "アルコ-ル耐性の個人差" 病態生理. 9. 948-955 (1990)

    • Related Report
      1990 Annual Research Report
  • [Publications] 原田 勝二: "エタノ-ルおよびアルデヒド代謝の人種的遺伝的要因" 医学のあゆみ. 154. 817-822 (1990)

    • Related Report
      1990 Annual Research Report
  • [Publications] 原田 勝二: "Glutathion SーTransferase 1 gene deletionおよびApolipoprotein Bー100 gene 3^'hypervariable regionのPCR法による解析とその応用" 2. 132-137 (1990)

    • Related Report
      1990 Annual Research Report
  • [Publications] 針原伸二: "ミトコンドリアDNA多型の家系調査" 日本法医学雑誌. 43. 257-261 (1989)

    • Related Report
      1989 Annual Research Report
  • [Publications] Keiko Washio: "Prob walking:development of novel probes for DNA fingerprinting" Human Genetics. 83. 223-226 (1989)

    • Related Report
      1989 Annual Research Report
  • [Publications] Keiko Washio: "Individual Identification of Non-human Primates Using DNA Fingerprinting" Primates. 30. 217-221 (1989)

    • Related Report
      1989 Annual Research Report
  • [Publications] Keiko Washio: "Effects of Cytosine Methylation on DNA Typing" J.Forensic Sciences. 35. (1990)

    • Related Report
      1989 Annual Research Report
  • [Publications] 本田克也: "硬組織から抽出したDNAによる性別判定" 日本法医学雑誌.

    • Related Report
      1989 Annual Research Report

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Published: 1989-04-01   Modified: 2016-04-21  

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