| 摘要: |
| 【目的】研究猪肌肉组织中猪载脂蛋白E(Apolipoprotein E,ApoE基因 5′调控区的DNA甲基化状况。【方法】以180日龄长白母猪为试验动物，采集其肌肉组织，提取其DNA，经亚硫酸氢盐修饰后用作PCR反应模板;通过生物信息学预测猪ApoE5′调控区潜在的CpG岛,根据预测结果选用甲基化特异性引物对CpG岛进行PCR扩增，对回收、纯化的PCR产物进行克隆和测序，并与GenBank中的ApoE)5′调控区序列进行比对，分析CpG岛的甲基化水平。【结果】生物信息学预测发现，猪ApoE基因5′ 调控区有2个CpG 岛，第1个CpG岛有10个CpG位点，第2个CpG岛有12个CpG位点；PCR扩增获得长度分别为233 bp的CpG 岛1和288 bp的CpG 岛2 序列，这2个CpG 岛均存在着9个甲基化CpG位点。【结论】猪ApoE基因5′调控区是CpG 位点的富集区域，2个CpG岛存在不同程度的甲基化。 |
| 关键词: 猪 肌肉组织 ApoE基因；DNA甲基化；CpG岛 |
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| 基金项目:广东省自然科学基金项目(10451022501005709)；国家生猪现代产业技术体系建设专项(CARS-36)；广东省科技计划项目(B1101328) |
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| DNA methylation in the 5′ regulatory region of porcineApoEin muscle tissues |
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LI Shixin, LI Jiaqi, TIAN Yunbo,et al
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Zhongkai University of Agriculture and Engineering,et al
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| Abstract: |
| 【Objective】This study focused on DNA methylation levels of the 5′regulatory region of porcine ApoEgene in muscle tissues of Landrace sows.【Method】Landrace sows at the age of 180 days were used as experimental animals,and muscle tissues were collected for subsequent DNA extraction.Modified DNA using sodium bisulfite was employed as the PCR reaction templates. The potential CpG islands in the porcineApoE5′regulatory region were predicted using bioinformatics and methylation-specific primers obtained from the prediction were chosen for amplifying the potential CpG islands. PCR amplicons were recycled,purified,and subsequently cloned into pMD-18T vectors for sequencing and then the sequences were aligned with that of the porcine ApoE5′regulatory region from GenBank for analyzing the methylation levels of CpG islands in this region.【Result】The results showed that two CpG islands within the porcine ApoEgene 5′regulatory region were found using bioinformatics and there were ten CpG sites within CpG island 1 and twelve within CpG island 2. Two sequences including the 233 bp length of CpG island 1 and the 288 bp of CpG island 2 were obtained. There were nine methylated sites within the two CpG islands.【Conclusion】The CpG sites were rich in the 5′regulatory region of porcineApoEgene and the two CpG islands were methylated in different degrees. |
| Key words: pig muscle tissue ApoEgene DNA methylation CpG island |
