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Development and application of a method to study DNA methylation and chromatin structure at the chromosome domain level

Research Project

Project/Area Number 06680669
Research Category

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

Allocation TypeSingle-year Grants
Research Field Molecular biology
Research InstitutionKyushu University

Principal Investigator

SASAKI Hiroyuki  Kyushu University, Institute of Genetic Information, Associate Professor, 遺伝情報実験施設, 助教授 (30183825)

Project Period (FY) 1994 – 1995
Project Status Completed (Fiscal Year 1995)
Budget Amount *help
¥2,300,000 (Direct Cost: ¥2,300,000)
Fiscal Year 1995: ¥1,300,000 (Direct Cost: ¥1,300,000)
Fiscal Year 1994: ¥1,000,000 (Direct Cost: ¥1,000,000)
KeywordsDNA methylation / Chromatin / CpG island
Research Abstract

In order to develop a method to study DNA methylation and chromtin structure of large genomic regions, we combined the methylation-sensitive restriction assay and nuclease-sensitivity assay with field-inversion gel electrophoresis (FIGE). The distal region of mouse chromosome 7, which comprises the imprinted ins2/lgf2/H19 genes, was analyzed in model expeniments. When 100 kb of the region was scanned by this method for DNasel hypersensitive sites, three new clusters of hypersensitive sites were identified. It was possible to study fragments larger than 25 kb. Similarly, it was possible to determine the methylation status of genomic regions larger than 25 kb after partial digestion with HpaII and HhaI.The reliability of these methods was comfirmed by examining the same regions by the conventional, short-range methods. We also found that partial degestion with HpaII and HhaI was useful for identifying and mapping CpG islands in native genomic and cloned DNA.The usefulness of the method has been shown with lambda, cosmid and P1 clones containing various inserts and a fragment as large as 85 kb could be analyzed at once. These methods are quite simple and reliable and thus can be applied to the long-range analysis of genome and chromatin structure.

Report

(3 results)
  • 1995 Annual Research Report   Final Research Report Summary
  • 1994 Annual Research Report
  • Research Products

    (6 results)

All Other

All Publications (6 results)

  • [Publications] Sasaki, H. et al.: "Temporal and spatial regulation of H19 imprinting in normal and uniparental embryos." Development. 121. 4195-4202 (1995)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1995 Final Research Report Summary
  • [Publications] Surani, M. A. et al.: "Genomic imprinting : causes and consequences" Cambridge University Press (eds. Ohlsson, R., Hall, K. & Ritzen, M.), 374 (1995)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1995 Final Research Report Summary
  • [Publications] Sasaki, H.et al.: "Temporal and spatial regulation of H19 imprinting in normal and uniparental embryos." Development. 121. 4195-4202 (1995)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1995 Final Research Report Summary
  • [Publications] Surani, M.A.et al.: "Imprinting of H19 and Xist in uniparental embryos. In : Genomic imprinting : causesand consequences." Cambridge University Press (eds.Ohlsson, R., Hall, K.& Ritzen, M.). 142-156 (1995)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1995 Final Research Report Summary
  • [Publications] Sasaki, H. et al.: "Temporal and spatial regulation of H19 imprinting in normal and uniparental embryos." Development. 121. 4195-4202 (1995)

    • Related Report
      1995 Annual Research Report
  • [Publications] Surani, M. A. et al.: "Genomic imprinting : causes and consequences" Cambridge University Press (eds. Ohlsson, R., Hall, K. & Ritzen, M.), 374 (1995)

    • Related Report
      1995 Annual Research Report

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Published: 1994-04-01   Modified: 2016-04-21  

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