MyoD1启动子区甲基化及mRNA表达分析  

MyOD1 promoter region methylation and mRNA expression analysis

作　　者：李雄[1,2] 田念念 宋林锦 陈晨 许厚强[1,2] LI Xiong;TIAN Nian-nian;SNG Lin-jin;CHEN Chen;XU Hou-qiang(Key Laboratory of Animal Genetics,Breeding and Reproduction in the Plateau Mountains Region of Ministry of Education,Guizhou University/Key Laboratory of Animal Genetics,Breeding and Reproduce in Guizhou Province,Guiyang 550025,China;College of Animal Sciences,Guizhou University,Guiyang 550025,China)

机构地区：[1]高原山地动物遗传育种与繁殖教育部重点实验室/贵州省动物遗传育种与繁殖重点实验室,贵州贵阳550025 [2]贵州大学动物科学学院,贵州贵阳550025

出　　处：《生物技术》2021年第2期112-120,159,共10页Biotechnology

基　　金：国家科技支撑计划项目(2015BAD03B02-3);贵州省农业重大产业科学研究攻关项目(黔教合KY字[2019]011)。

摘　　要：[目的]检测关岭牛背最长肌中MyoD1基因甲基化水平及mRNA表达量,分析胎牛与成年牛甲基化水平差异。[方法]以关岭牛胎牛以及成年牛(24月龄)为研究对象,利用亚硫酸氢盐介导的未甲基化胞嘧啶向尿嘧啶的转化以及用高通量测序方法检测其背最长肌肉中的甲基化表达水平;通过qRT-PCR检测MyoD1、DNMT1基因在关岭牛胎牛背最长肌中中的表达量。[结果]胎牛背最长肌中MyoD1启动子甲基化率为15.4%、1.4%、0.8%,成年牛为20%、1.1%、1.2%,且胎牛中MyoD1启动子甲基化水平极显著低于成年牛(P <0.01);通过甲基检测结果筛选出甲基化位点CpG-1-2相关转录因子结合位点ZNF449;MyoD1在关岭牛胎牛背最长肌中的表达量极显著高于成年牛(P <0.01),DNMT1基因在关岭牛胎牛背最长肌中的表达量极显著低于成年牛(P <0.01)。[结论]MyoD1启动子甲基化水平在关岭牛胎牛、成年牛存在差异,DNMT1的表达与甲基化水平一致,预测启动子区可能调控的甲基化位点。推测MyoD1启动子甲基化在肌肉的生长发育过程中有重要的作用,为进一步研究关岭牛MyoD1启动子甲基化提供理论基础。[Objective]To detect the methylation level and mRNA expression of MYOD1 gene in longissimus dorsi muscle of Guanling cattle,and to analyze the difference of methylation level between fetal cattle and adult cattle.[Method]Bisulfite mediated transformation of unmethylated cytosine was used to uracil and high-throughput sequencing to detect the expression of methylation in the longissimus dorsi muscle of Guanling bovine(24 months old). The expression levels of MyOD1 and DNMT1 genes in longissimus dorsi muscle of Guanling cattle were detected by qRT-PCR. [Result]The methylation rates of MyOD1 promoter in longissimus dorsi muscle of fetal cattle were 15. 4%,1. 4%,0. 8%,and those of adult cattle were 20%,1. 1%,1.2%,and the methylation level of MyOD1 promoter in fetal cattle was significantly lower than that in adult cattle(P < 0. 01);The methylation site CpG-1-2 related transcription factor binding site ZNF449 was screened out by methyl detection results;The expression level of MyOD1 in longissimus dorsi muscle of Guanling fetal bovine was significantly higher than that of adult bovine(P < 0. 01),and the expression level of DNMT1 in longissimus dorsi muscle of Guanling fetal bovine was significantly lower than that of adult bovine(P < 0. 01). [Conclusion]The methylation level of MyOD1 promoter is different between fetal and adult Guanling cattle,and the expression of DNMT1 is consistent with the methylation level,which predicts the methylation sites that may be regulated by the promoter region. It is speculated that MyOD1 promoter methylation plays an important role in muscle growth and development,and provides a theoretical basis for further study of MyOD1 promoter methylation in Guanling cattle.

关 键 词：关岭牛 高通量检测 DNA甲基化 转录因子 

分 类 号：S823.81[农业科学—畜牧学]