De novo DNA methylation of the 35S enhancer revealed by high-resolution methylation analysis of an entire T-DNA segment in transgenic gentian
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概要
- 論文の詳細を見る
We have found that cauliflower mosaic virus (CaMV) 35S promoter-specific transgene silencing is mediated by DNA methylation in gentian (Gentiana triflora × G. scabra). De novo methylation of asymmetric cytosines (CpHpH; where H is A, C, or T) sequence has been detected at the enhancer region (−148 to −85) of the 35S promoter in transgenic gentians, and is thought to be responsible for the silencing mechanism. To clarify the concept of de novo methylation, the present study examined the detailed DNA methylation profile of the entire T-DNA sequence (ca. 4 kb) integrated into transgenic gentians. Although highly methylated cytosines at CpG and CpWpG (W is A or T) sequences were broadly distributed, except in the sGFP coding region, highly methylated cytosines at CpHpH and CpCpG sequences were mainly limited to the 35S enhancer region. In addition to the previously identified de novo methylation peak (−148 to −85), another peak was discovered at −298 to −241. Electrophoretic mobility shift assays showed that gentian nuclear extracts could bind to the corresponding probes (−149 to −124 and −275 to −250), and that the probes could compete with one another for binding. Thus, a nuclear factor might be involved in the de novo methylation of the two regions. In addition, the present data indicated that the methylation patterns at CpCpG sites could be categorized as CpHpH methylation rather than CpWpG methylation.
- 2011-03-25
著者
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箕作 和彦
大阪府立大学院生命環境科学研究科
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小田 雅行
大阪府立大学大院生命環境科学研究科
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Oda Masayuki
Graduate School Of Life And Environmental Sciences Osaka Prefecture University
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Koizumi Nozomu
Graduate School Of Agricultural Sciences Osaka Prefectural University
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淨閑 正史
大阪府立大院生命環境科学研究科
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YAMASAKI Satoshi
Graduate School of Life and Environmental Sciences, Osaka Prefecture University
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小田 雅行
大阪府立大学大学院生命環境科学研究科
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YAMAMURA Saburo
Iwate Biotechnology Research Center
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Johkan Masafumi
Graduate School Of Life And Environmental Sciences Osaka Prefecture University
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Mitsukuri Kazuhiko
Graduate School Of Life And Environmental Sciences Osaka Prefecture University
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Mishiba Kei-ichiro
Graduate School Of Life And Environmental Sciences Osaka Prefecture University
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Mishiba Keiichirou
Graduate School Of Life And Environmental Sciences Osaka Prefecture University
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Nishihara Masahiro
Iwate Biotechnology Research Center
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TSUKATANI Nobue
Iwate Biotechnology Research Center
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Nakata Masaru
Bioproduction Res. Inst. National Inst. Of Advanced Industrial Sci. And Technol. (aist)
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Nakatsuka Takashi
Iwate Biotechnology Res. Center
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Nishihara Masahiro
Iwate Biotechnology Res. Center
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Johkan Masafumi
Graduate School Of Life And Environmental Sciences Osaka Prefecture Univ.
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Nishihara Masahiro
Bioproduction Res. Inst. National Inst. Of Advanced Industrial Sci. And Technol. (aist)
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Oda Masayuki
Graduate School Of Agriculture And Life Science The University Of Tokyo
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Yamasaki Satoshi
Graduate School Of Life And Environmental Sciences Osaka Prefecture Univ.
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Yamasaki Satoshi
Graduate School Of Life And Environmental Sciences Osaka Prefecture University
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