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

发布时间：2018-03-15 21:40

本文选题：鸭　切入点：肌细胞生成素　出处：《湖南农业大学学报(自然科学版)》2017年02期 　论文类型：期刊论文

【摘要】：采用RT PCR技术扩增和克隆鸭Myo G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 Mass Array技术检测Cp G岛在鸭肌肉组织中的甲基化水平,用q RT PCR检测Myo G基因的表达量。结果表明,扩增得到鸭Myo G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Cp G岛,其中Cp G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;Cp G位点甲基化频率存在个体差异,22%Cp G位点的甲基化频率与Myo G的m RNA表达量呈负相关(P0.05),78%Cp G位点的甲基化频率呈正相关(P0.05),其中,腿肌甲基化位点Cp G_1、Cp G_26.27.28.29的甲基化频率与Myo G基因表达水平均呈显著相关(P0.05)。Myo G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Myo G基因转录的潜在甲基化位点,其中Cp G_1与Cp G_26.27.28.29能通过DNA甲基化修饰影响Myo G基因在鸭腿肌中的转录。本研究结果可为鸭Myo G基因转录调控提供参考依据。
[Abstract]:The promoter of duck Myo G gene was amplified and cloned by RT PCR, and its promoter sequence was analyzed by bioinformatics. The methylation level of CP G island in duck muscle tissue was detected by Sequenom Mass Array technique. Q RT PCR was used to detect the expression of Myo G gene. The results showed that the promoter sequence of duck Myo G gene was 2 730 bp. after predicting the promoter sequence, two CpG islands were found. There are 5 transcription factor binding sites and several eukaryote structural elements in CpG island (Cp2 536 ~ 1 997 BP). Promoter methylation rate was not clustered together. There were individual differences in methylation frequency of CP G locus. There was a negative correlation between methylation frequency at CP G site and m RNA expression of Myo G. There was a positive correlation between methylation frequency at CpG site and m RNA expression level in Myo G locus, and there was a positive correlation between methylation frequency at CpG locus and m RNA expression level in Myo G locus. The methylation frequency of CP G 26.27.28.29 in leg muscle methylation site was significantly correlated with the expression level of Myo G gene. There were significant differences in the transcriptional regulation mechanism between duck and mammal. Potential methylation sites for transcription, CpG1 and CP G26.27.28.29 can affect the transcription of Myo G gene in duck leg muscle by DNA methylation. The results can provide a reference for the regulation of Myo G gene transcription in duck leg muscle.
【作者单位】：四川农业大学动物遗传育种研究所;
【基金】：国家自然科学基金项目(31301964) 四川省科学技术厅基金项目(2015JY0110)
【分类号】：Q78;S834

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