Analysis of DNA methylation in various swine tissues.
Analysis of DNA methylation in various swine tissues.
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DOI：
10.1371/journal.pone.0016229
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发表时间：
2011-01-21
期刊：
影响因子：
3.7
通讯作者：
Zhao Z
中科院分区：
文献类型：
--
作者：
Yang C;Zhang M;Niu W;Yang R;Zhang Y;Qiu Z;Sun B;Zhao Z
DNA甲基化在真核生物的生物发育和组织分化过程中对基因表达起着重要的调控作用。本研究采用荧光标记甲基化敏感扩增多态性（F-MSAP）方法，对莱芜猪肌肉、心、肝、脾、肺、肾和胃中胞嘧啶甲基化的程度和模式进行了分析，并对这7种组织中的特异性甲基化模式进行了检测。使用16对选择性引物，总共扩增了96，371个片段，每个片段代表由EcoRI + Hpa II和EcoRI + Msp I中的一个或两个切割的识别位点，Hpa II和Msp I是同源酶。在7种组织中，共发现50，094个位点在胞嘧啶处甲基化。差异消化率显示，肌肉、心脏、肝脏、脾脏、肺脏、肾脏和胃的DNA甲基化发生率分别为53.99%、51.24%、50.18%、53.31%、51.97%、51.15%和53.39%。此外，DNA甲基化水平的差异意味着这种变化可能与组织分化，生长和发育过程中的特定基因表达有关。F-MSAP图谱中出现3种类型的条带，这3种类型的条带在7种组织中的总数分别为46，277、24，801和25，293条。此外，在猪的7种组织中发现了不同的甲基化模式，并且几乎所有F-MSAP检测到的甲基化模式都可以通过Southern分析得到证实。结果清楚地表明，F-MSAP技术可以适用于在猪基因组中的大规模DNA甲基化检测。
DNA methylation is known to play an important role in regulating gene expression during biological development and tissue differentiation in eukaryotes. In this study, we used the fluorescence-labeled methylation-sensitive amplified polymorphism (F-MSAP) method to assess the extent and pattern of cytosine methylation in muscle, heart, liver, spleen, lung, kidney and stomach from the swine strain Laiwu, and we also examined specific methylation patterns in the seven tissues. In total, 96,371 fragments, each representing a recognition site cleaved by either or both EcoRI + HpaII and EcoRI + MspI, the HpaII and MspI are isoschizomeric enzymes, were amplified using 16 pairs of selective primers. A total of 50,094 sites were found to be methylated at cytosines in seven tissues. The incidence of DNA methylation was approximately 53.99% in muscle, 51.24% in the heart, 50.18% in the liver, 53.31% in the spleen, 51.97% in the lung, 51.15% in the kidney and 53.39% in the stomach, as revealed by the incidence of differential digestion. Additionally, differences in DNA methylation levels imply that such variations may be related to specific gene expression during tissue differentiation, growth and development. Three types of bands were generated in the F-MSAP profile, the total numbers of these three types of bands in the seven tissues were 46,277, 24,801 and 25,293, respectively. In addition, different methylation patterns were observed in seven tissues from pig, and almost all of the methylation patterns detected by F-MSAP could be confirmed by Southern analysis using the isolated amplified fragments as probes. The results clearly demonstrated that the F-MSAP technique can be adapted for use in large-scale DNA methylation detection in the pig genome.
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