摘要
为研究鸡脂肪组织miR-17-92基因簇宿主基因(MIR17HG)启动子的甲基化情况,实验以东北农业大学高、低腹脂双向选择品系肉鸡为实验材料,采用Sequenom MassARRAY法检测鸡腹部脂肪组织MIR17HG启动子区CpG岛的甲基化;利用CpG甲基转移酶处理MIR17HG启动子片段,采用报告基因技术检测DNA甲基化对MIR17HG启动子活性的影响。结果表明:高、低脂鸡脂肪组织中MIR17HG启动子区都存在一定程度的甲基化,其中2周龄低脂鸡脂肪组织MIR17HG启动子区的整体甲基化程度显著高于高脂鸡(P<0.05),4个CpG位点(25、26、27、28)的甲基化在高、低脂鸡脂肪组织间差异均显著(P<0.05);3周龄高、低脂鸡脂肪组织中MIR17HG启动子区的甲基化程度无显著差异(P>0.05);7周龄低脂鸡脂肪组织MIR17HG启动子的2个CpG位点(41和42)的甲基化程度显著低于高脂鸡(P<0.05)。总体看,高、低脂鸡脂肪组织MIR17HG的启动子甲基化随年龄增长呈下降趋势;体外甲基化分析发现,DNA甲基化抑制MIR17HG启动子活性。
The aim of this study was to investigate DNA methylation of the promoter region of chicken miR-17-92 cluster host gene(MIR17HG).In this study,promoter methylation of MIR17HG in adipose tissue of the broilers from the Northeast Agricultural University(NEAU)broiler lines divergently selected for abdominal fat content(NEAUHLF)was detected by Sequenom Mass ARRAY.In addition,chicken MIR17HG promoter fragment was methylated in vitro by CpG methyltransferase,the effect of DNA methylation on the promoter activity was assessed by reporter gene assay.The results showed,at two weeks of age,the promoter methylation level of MIR17HG in adipose tissue was significantly higher in lean line than in fat line,and the methylation levels of CpG site 25,CpG site 26,CpG site 27 and CpG site 28 were significantly different(P<0.05)between the lean and fat lines.At three weeks of age,no difference in the overall promoter methylation level of MIR17HG in adipose tissue was observed between the lean and fat lines(P>0.05).At seven weeks of age,the overall promoter methylation levels of MIR17HG in adipose tissue in both lean and fat lines were similar,but the methylation levels of CpG site 41 and CpG site 42 were significantly lower in lean line than in fat line(P<0.05).On the whole,the methylation levels of chicken MIR17HG promoter in both chicken lines increased with age.Reporter assay showed that DNA methylation inhibited MIR17HG promoter activity.
作者
邢天宇
崔婷婷
黄佳新
闫晓红
李辉
王宁
XING Tian-yu;CUI Ting-ting;HUANG Jia-xin;YAN Xiao-hong;LI Hui;WANG Ning(Key Laboratory of Chicken Genetics and Breeding,Ministry of Agriculture,Key Laboratory of Animal Genetics,Breeding and Reproduction,Education Department of Heilongjiang Province,College of Animal Science and Technology,Northeast Agricultural University,Heilongjiang Harbin 150030,China)
出处
《中国畜牧杂志》
CAS
北大核心
2018年第11期49-54,共6页
Chinese Journal of Animal Science
基金
国家自然科学基金项目(31372299)。
