DNA of Drosophila melanogaster contains 5-methylcytosine
DNA of Drosophila melanogaster contains 5-methylcytosine
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DOI：
10.1093/emboj/19.24.6918
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发表时间：
2000-12-15
期刊：
影响因子：
11.4
通讯作者：
Jeltsch, A
中科院分区：
文献类型：
--
作者：
Gowher, H;Leismann, O;Jeltsch, A
人们普遍认为，果蝇的 DNA 不含 5-甲基胞嘧啶，而 5-甲基胞嘧啶对于大多数真核生物的发育至关重要。我们开发了一种新的、高度特异性和灵敏的检测方法来检测基因组 DNA 中 5-甲基胞嘧啶的存在。 DNA 被降解为核苷，通过 HPLC 纯化为 5-甲基胞嘧啶，并使用脱氧核苷激酶和 [gamma-P-32]ATP 进行放射性标记，以通过 1D-和 2D-TLC 进行检测。使用该测定，我们在此表明，黑腹果蝇 DNA 中 5-甲基胞嘧啶的水平与成年果蝇中 1000-2000 个胞嘧啶残基中的 1 相似。 DNA 甲基化在黑腹果蝇发育的所有阶段均可检测到。
It is commonly accepted that the DNA of Drosophila melanogaster does not contain 5-methylcytosine, which is essential in the development of most eukaryotes. We have developed a new, highly specific and sensitive assay to detect the presence of 5-methylcytosine in genomic DNA. The DNA is degraded to nucleosides, 5-methylcytosine purified by HPLC and, for detection by 1D- and 2D-TLC, radiolabeled using deoxynucleoside kinase and [gamma-P-32]ATP. Using this assay, we show here that 5-methylcytosine occurs in the DNA of D.melanogaster at a level of similar to1 in 1000-2000 cytosine residues in adult flies. DNA methylation is detectable in all stages of D.melanogaster development.