鸡脂肪组织TCF 21基因启动子区DNA甲基化与其表达的关系  

The Relationship between Promoter Region DNA Methylation of TCF 21 Gene and Its Expression in Chicken Adipose Tissue

作　　者：刘雨萌 马艳艳 姜海煦 张心扬 武春艳 程博涵[1,2,3] 李辉 LIU Yumeng;MA Yanyan;JIANG Haixu;ZHANG Xinyang;WU Chunyan;CHENG Bohan;LI Hui(Key Laboratory of Chicken Genetics and Breeding,Ministry of Agriculture and Rural Affairs,Harbin 150030,China;Key Laboratory of Animal Genetics,Breeding and Reproduction,Education Department of Heilongjiang Province,Harbin 150030,China;College of Animal Sciences and Technology,Northeast Agricultural University,Harbin 150030,China)

机构地区：[1]农业农村部鸡遗传育种重点实验室,哈尔滨150030 [2]黑龙江普通高等学校动物遗传育种与繁殖重点实验室,哈尔滨150030 [3]东北农业大学动物科学技术学院,哈尔滨150030

出　　处：《畜牧兽医学报》2021年第12期3375-3389,共15页ACTA VETERINARIA ET ZOOTECHNICA SINICA

基　　金：国家自然科学基金(31772586);财政部和农业农村部:国家现代农业产业技术体系(CARS-41)。

摘　　要：旨在研究鸡脂肪组织中TCF 21基因启动子区DNA甲基化水平与其表达的关系。以东北农业大学高、低腹脂双向选择品系(简称高、低脂系)第24世代7周龄肉鸡为试验材料,利用RT-qPCR检测高、低脂系肉鸡腹部脂肪组织中TCF 21基因的mRNA表达水平;利用生物信息学和双荧光素酶报告系统分析TCF 21基因启动子的结构与功能;利用Sequenom MassARRAY飞行质谱检测高、低脂系肉鸡腹部脂肪组织中TCF 21基因启动子区CpG位点的甲基化水平;利用CpG甲基转移酶处理TCF 21启动子报告基因质粒,分析DNA甲基化对TCF 21基因启动子活性的影响。结果显示,高脂系肉鸡腹部脂肪组织中TCF 21基因的mRNA表达水平极显著高于低脂系(P<0.001);TCF 21基因的启动子区存在40个CpG位点,且在启动子的近端和远端均有分布,但不存在CpG岛;将TCF 21基因的启动子划分为5个功能区域,分别为R1区域(-2000~-1500 bp)、R2区域(-1500~-1000 bp)、R3区域(-1000~-500 bp)、R4区域(-500~-200 bp)和Core区域(-200~-100 bp);高脂系R2、R3和R2+R3区域的DNA甲基化水平显著或极显著高于低脂系(P<0.05或P<0.001);R2、R3、R2+R3区域的DNA甲基化水平与TCF 21基因mRNA表达水平呈显著正相关(R2区域:r=0.438,P<0.05;R3区域:r=0.371,P<0.05;R2+R3区域:r=0.489,P<0.05);R2区域的DNA甲基化显著抑制其转录活性(P<0.05)。综上所述,TCF 21基因在高、低脂系肉鸡腹部脂肪组织中的表达水平主要与其启动子R2区域的DNA甲基化水平有关。The aim of this study was to investigate the relationship between DNA methylation of TCF 21 gene promoter region and its expression in chicken adipose tissue.The mRNA expression levels of TCF 21 in abdominal adipose tissues of chickens which were at 7 weeks of age and from 24^(th) generation of Northeast Agricultural University broiler lines divergently selected for abdominal fat content(NEAUHLF)were detected by RT-qPCR.The structure and function of TCF 21 promoter were analyzed by bioinformatics and luciferase reporter system.The methylation level of CpG sites in the promoter region of TCF 21 in the abdominal adipose tissues of the fat and lean broilers was detected by Sequenom MassARRAY platform.The TCF 21 promoter reporter gene plasmid was treated with CpG methyltransferase to analyze the effect of DNA methylation on the activity of TCF 21 promoter.The results showed that the mRNA expression levels of TCF 21 in the abdominal adipose tissue of fat broilers were extremely significantly higher than those of lean broilers(P<0.001).There were 40 CpG sites in the promoter region,which were distributed in the proximal and distal parts of the promoter,but there was no CpG island in the promoter region of chicken TCF 21 gene.The promoter of TCF 21 gene was divided into 5 functional regions,namely R1 region(-2000~-1500 bp),R2 region(-1500~-1000 bp),R3 region(-1000~-500 bp),R4 region(-500~-200 bp)and Core region(-200~-100 bp),respectively.The DNA methylation levels in the R2,R3 and R2+R3 regions of TCF 21 promoter in the fat broilers were significantly or extremely significantly higher than those of the lean broilers(P<0.05 or P<0.001).The DNA methylation levels in the R2,R3 and R2+R3 regions of TCF 21 promoter were significantly positively correlated with TCF 21 mRNA expression levels(R2 region:r=0.438,P<0.05;R3 region:r=0.371,P<0.05;R2+R3 region:r=0.489,P<0.05).DNA methylation significantly inhibited the transcriptional activity of R2 region of TCF 21 promoter(P<0.05).In summary,the expression level of TCF 21 gene in a

关 键 词：鸡 TCF21基因 启动子 DNA甲基化 基因表达 脂肪组织 

分 类 号：S831[农业科学—畜牧学]