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  1. 原著論文

Methylated site display (MSD)-AFLP, a sensitive and affordable method for analysis of CpG methylation profiles

https://repo.qst.go.jp/records/47729
https://repo.qst.go.jp/records/47729
c6729ca3-8812-4eb6-bf7a-d5e2fb53b97c
Item type 学術雑誌論文 / Journal Article(1)
公開日 2017-04-20
タイトル
タイトル Methylated site display (MSD)-AFLP, a sensitive and affordable method for analysis of CpG methylation profiles
言語
言語 eng
資源タイプ
資源タイプ識別子 http://purl.org/coar/resource_type/c_6501
資源タイプ journal article
アクセス権
アクセス権 metadata only access
アクセス権URI http://purl.org/coar/access_right/c_14cb
著者 Aiba, Toshiki

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WEKO 478912

Aiba, Toshiki

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Saito, Toshiyuki

× Saito, Toshiyuki

WEKO 478913

Saito, Toshiyuki

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Hayashi, Akiko

× Hayashi, Akiko

WEKO 478914

Hayashi, Akiko

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Sato, Shinji

× Sato, Shinji

WEKO 478915

Sato, Shinji

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Yunokawa, Harunobu

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WEKO 478916

Yunokawa, Harunobu

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Maruyama, Toru

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WEKO 478917

Maruyama, Toru

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Fujibuchi, Wataru

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WEKO 478918

Fujibuchi, Wataru

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Kurita, Hisaka

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WEKO 478919

Kurita, Hisaka

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Tohyama, Chiharu

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WEKO 478920

Tohyama, Chiharu

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Ohsako, Seiichiroh

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WEKO 478921

Ohsako, Seiichiroh

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相場 俊樹

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WEKO 478922

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齋藤 俊行

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WEKO 478923

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林 昭子

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WEKO 478924

en 林 昭子

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抄録
内容記述タイプ Abstract
内容記述 Background: It has been pointed out that environmental factors or chemicals can cause diseases that are developmental
in origin. To detect abnormal epigenetic alterations in DNA methylation, convenient and cost-effective
methods are required for such research, in which multiple samples are processed simultaneously. We here present
methylated site display (MSD), a unique technique for the preparation of DNA libraries. By combining it with amplified
fragment length polymorphism (AFLP) analysis, we developed a new method, MSD-AFLP.
Results: Methylated site display libraries consist of only DNAs derived from DNA fragments that are CpG methylated
at the 5′ end in the original genomic DNA sample. To test the effectiveness of this method, CpG methylation levels in
liver, kidney, and hippocampal tissues of mice were compared to examine if MSD-AFLP can detect subtle differences
in the levels of tissue-specific differentially methylated CpGs. As a result, many CpG sites suspected to be tissue-specific
differentially methylated were detected. Nucleotide sequences adjacent to these methyl-CpG sites were identified
and we determined the methylation level by methylation-sensitive restriction endonuclease (MSRE)-PCR analysis
to confirm the accuracy of AFLP analysis. The differences of the methylation level among tissues were almost identical
among these methods. By MSD-AFLP analysis, we detected many CpGs showing less than 5% statistically significant
tissue-specific difference and less than 10% degree of variability. Additionally, MSD-AFLP analysis could be used to
identify CpG methylation sites in other organisms including humans.
Conclusion: MSD-AFLP analysis can potentially be used to measure slight changes in CpG methylation level. Regarding
the remarkable precision, sensitivity, and throughput of MSD-AFLP analysis studies, this method will be advantageous
in a variety of epigenetics-based research.
Keywords: DNA methylation profiling, AFLP, Epigenetics
書誌情報 BMC Molecular Biology

巻 18, 号 7, p. 1-11, 発行日 2017-03
出版者
出版者 BioMed Central
ISSN
収録物識別子タイプ ISSN
収録物識別子 1471-2199
DOI
識別子タイプ DOI
関連識別子 10.1186/s12867-017-0083-2
関連サイト
識別子タイプ URI
関連識別子 https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5345256/pdf/12867_2017_Article_83.pdf
関連名称 https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5345256/pdf/12867_2017_Article_83.pdf
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