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Establishment of carrier detection method of X-linked diseases by methylation-specific PCR

Research Project

Project/Area Number 11670752
Research Category

Grant-in-Aid for Scientific Research (C)

Allocation TypeSingle-year Grants
Section一般
Research Field Pediatrics
Research InstitutionNational Center of Neurology and Psychiatry (2000)
Shinshu University (1999)

Principal Investigator

KUBOTA Takeo  National Center of Neurology and Psychiatry, Head, 疾病研究第2部, 室長 (70293511)

Co-Investigator(Kenkyū-buntansha) AGEMATSU Kazunaga  Shinshu University School of Medicine, Assistant Professor, 大学院・医学研究科・移植免疫感染症, 講師 (60262721)
FUKUSHIMA Yoshimitsu  Shinshu University School of Medicine, Professor, 医学部・衛生学, 教授 (70273084)
GOTO Yu-ichi  National Center of Neurology and Psychiatry, Director, 疾病研究第2部, 部長 (20225668)
Project Period (FY) 1999 – 2000
Project Status Completed (Fiscal Year 2000)
Budget Amount *help
¥2,900,000 (Direct Cost: ¥2,900,000)
Fiscal Year 2000: ¥1,000,000 (Direct Cost: ¥1,000,000)
Fiscal Year 1999: ¥1,900,000 (Direct Cost: ¥1,900,000)
KeywordsX-chromosome / inactivation / methylation / carrier detection / methylation-specific PCR / androgen receptor gene / HUMARA gene / nonrandom pattern / メチレーション
Research Abstract

The pattern of X-chromosome inactivation in females has currently been evaluated by the assays through diffential methylation in the genes between the active and the inactive X chromosomes using methylation-sensitive enzymes. We report a new assay in the human androgen receptor (HUMARA) locus using a methylation-specific PCR (M-PCR) technique, independent of the use of restriction enzymes. The assay involves the chemical modification of DNA with sodium bisulfite and the subsequent PCR.By the assay with specific primers for the methylated allele, we obtained an X-inactivation pattern based on the ratio of the material inactive X to the paternal inactive X.These patterns were consistent with those by the conventional PCR assay at the same locus in 48 female cases. We also obtained another X-inactivation pattern based on the ratio of the maternal active X to the paternal active X using specific primers for the unmethylated allele. The latter pattern were complementary to the former pattern, and combination of these patterns produced a reliable X-inactivation pattern. By the assay, twelve (11%) of the 105 normal females showed nonrandom inactivation patterns (>80 : 20 or <20 : 80). Four patients with a X ; autosome translocation showed extremely nonrandom patterns, and these results were consistent with those by the previous molecular/cytogenetic studies. We conclude that M-PCR provides a accurate assay for X-inactivation, which can be performed on various DNA samples unsuitable for restriction digestion.

Report

(3 results)
  • 2000 Annual Research Report   Final Research Report Summary
  • 1999 Annual Research Report
  • Research Products

    (20 results)

All Other

All Publications (20 results)

  • [Publications] Kubota T, et al.: "A new assay for the analysis of X-chromosome inactivation..."Hum Genet. 104. 49-55 (1999)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] Kubota T, et al.: "Borjeson-Forssmann-Lehmann syndrome in a womoan with skewed X-inactivation..."Am J Med Genet. 87. 258-261 (1999)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] Wadat,Kubota T, et al.: "Molecular genetic study of Japanese patients with X-linked..."Am J Med Genet. 94. 242-248 (2000)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] Nakane T,Kubota T, et al.: "Opiz Trigonocephaly(C)-like syndrome,or Bohring-Opitz syndrome"Am J Med Genet. 92. 361-362 (2000)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] Seki H,Kubota T,Ikegawa S, et al.: "Mutation frequencies of EXT1 and EXT2 in 43 Japanese families..."Am J Med Genet. 99. 59-62 (2001)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] Kubota T, Nonoyama S, Tonoki H, Masuno M, Imaizumi K, Kojima M, Wakui K, Shimadzu M, Fukushima Y: "A new assay for the analysis of X-choromosome inactivation based on methylation-specific PCR."Hum Genet. 104. 49-55 (1999)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] Nakajima K, Sakurai A, Kubota T, Katai M, Mori J, Aizawa T, Fukushima Y, Hashizume K: "Multiple Endocrine Neoplasia Type 1 Concomitant with Prader-Willi Syndrome : Case Report and Genetic Diagnosis."Am J Med Sci. 317. 346-349 (1999)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] Kubota T, Oga S, Ohashi H, Iwamoto Y, Fukushima Y: "Borjeson-Forssman-Lehmann syndrome in a woman with skewed X-chromosome inactivation."Am J Med Genet. 87. 258-261 (1999)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] Nishimura G, Kuwashima S, Kohno T, Teramoto C, Hiroshi W, Kubota T: "Fetal polycystic kidney disease in oro-facio-digital syndrome type I."Pediatr Radiol. 29. 506-508 (1999)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] Nakane T, Kubota T, Fukushima F, Hata Y, Ishii J, Komiyama A: "Opiz Trigonocephaly (C)-like syndrome, or Bohring-Opitz syndrome : Another example."Am J Med Genet. 92. 361-362 (2000)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] Wada t, Kubota T, Fukushima Y, Saitoh S.: "Molecular genetic study of Japanese patients with X-linked -thalassemia/mental retardation syndrome (ATR-X)."Am J Med Gene. 94. 242-248 (2000)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] Seki H, Kubota T, Ikegawa S, Haga N, Fujioka F, Ohzeki S, Wakui K, Yoshikawa H, Takaoka K, Fukushima Y: "Mutation frequencies of EXT1 and EXT2 in 43 Japanese families with hereditary multiple exostoses."Am J Med Gene. 99. 59-62 (2001)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] Kubota T, et al.: "A new assay for the analysis of X-chromosome inactivation..."Hum Genet. 104. 49-55 (1999)

    • Related Report
      2000 Annual Research Report
  • [Publications] Kubota T, et al.: "Borjeson-Forssmann-Lehmann syndrome in a womoan with skewed X-inactivation..."Am J Med Genet. 87. 258-261 (1999)

    • Related Report
      2000 Annual Research Report
  • [Publications] Wada t,Kubota T, et al..: "Molecular genetic study of Japanese patients with X-linked..."Am J Med Genet. 94. 242-248 (2000)

    • Related Report
      2000 Annual Research Report
  • [Publications] Nakane T.Kubota T, et al.: "Opiz Trigonocephaly (C)-like syndrome, or Bohring-Opitz syndrome"Am J Med Genet. 92. 361-362 (2000)

    • Related Report
      2000 Annual Research Report
  • [Publications] Seki H,Kubota T,Ikegawa S, et al.: "Mutation frequencies of EXT1 and EXT2 in 43 Japanese families..."Am J Med Genet. 99. 59-62 (2001)

    • Related Report
      2000 Annual Research Report
  • [Publications] Kubota T, et al.: "A new assay for the analysis of X-chromosome inactivation based on the methylation specific PCR"Human Genetics. 104. 49-55 (1999)

    • Related Report
      1999 Annual Research Report
  • [Publications] Kubota T, et al.: "Borjeson-Forssman-Lehmann Syndrome in a woman with skewed X-chromosome inactivation"American Journal of Medical Genetics. 87. 258-261 (1999)

    • Related Report
      1999 Annual Research Report
  • [Publications] 久保田 健夫: "シリーズ最新医学講座-遺伝子診断 Technology編 Methylation-specific PCR(M-PCR)法"臨床検査. 43・12. 1533-1539 (1999)

    • Related Report
      1999 Annual Research Report

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Published: 1999-04-01   Modified: 2016-04-21  

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