鸭MyoG基因启动子的生物信息学分析及其甲基化频率对基因表达的影响
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国家自然科学基金项目(31301964);四川省科学技术厅基金项目(2015JY0110)


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    摘要:

    采用RT–PCR技术扩增和克隆鸭MyoG基因启动子,并对其启动子序列进行生物信息学分析,采用Sequenom MassArray技术检测CpG岛在鸭肌肉组织中的甲基化水平,用qRT–PCR检测MyoG基因的表达量。结果表明,扩增得到鸭MyoG基因启动子序列2 730 bp,对启动子序列预测后,发现存在2个CpG岛,其中CpG岛(–2 536~–1 997 bp)存在5个转录因子结合位点和多个真核生物结构元件。甲基化检测结果表明:在鸭的个体和组织水平上,启动子甲基化率均未聚类在一起;CpG位点甲基化频率存在个体差异,22% CpG位点的甲基化频率与MyoG的mRNA表达量呈负相关(P>0.05),78% CpG位点的甲基化频率呈正相关(P>0.05),其中,腿肌甲基化位点CpG_1、CpG_26.27.28.29的甲基化频率与MyoG基因表达水平均呈显著相关(P<0.05)。MyoG基因在鸭与在哺乳动物中的转录调控机制存在差异。试验中发现多个影响鸭MyoG基因转录的潜在甲基化位点,其中CpG_1与CpG_26.27.28.29能通过DNA甲基化修饰影响MyoG基因在鸭腿肌中的转录。本研究结果可为鸭MyoG基因转录调控提供参考依据。

    Abstract:

    Promoter sequence of gene MyoG in duck was amplified and cloned using RT–PCR, and its bioinformatic info was analyzed as well; the methylation level in CpG island (from –2 536 to –1 997 bp) of promoter MyoG in muscle tissues was detected using the Sequenom MassArray technique, meanwhile, its expression level was detected using qRT–PCR. The results showed that the amplified MyoG promoter sequences was 2 730 bp which contained 2 CpG islands in promoter region. The CpG island (from –2 536 to –1 997 bp) included 5 binding sites of transcription factor and several eukaryotes structure components. The methylation data showed that methylation status from different individuals and tissues did not clustered together. Methylation frequency in CpG loci varied with individuals, and the methylation frequency on 22% of locus were negatively correlated with MyoG expression levels (P>0.05), whereas, on 78% of locus were positively correlated (P>0.05) with that, among of them, they reached significant difference level on locus of CpG 1, CpG 26.27.28.29 from leg muscles (P<0.05). The mechanism of regulating transcription for MyoG was different from mammals. Among the potential locus influencing MyoG transcription, CpG_1 and CpG_26.27.28.29 might affect MyoG transcription in skeletal muscle tissues of duck leg through DNA methylation. This study might list foundation data for studying the regulatory mechanisms of MyoG transcription in duck.

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吴乾锋,刘贺贺,张涛,罗俊,胡博,王继文.鸭MyoG基因启动子的生物信息学分析及其甲基化频率对基因表达的影响[J].湖南农业大学学报:自然科学版,2017,43(2):.

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  • 在线发布日期: 2017-04-18
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