关岭牛MEF2C基因克隆与差异表达性分析  

Cloning and differential expression analysis of MEF2Cgene in Guanling cattle

作　　者：孙金魁 许厚强[1,2] 黄勇[1,2] 李永 熊讯 SUN Jinkui;XU Houqiang;HUANG Yong;LI Yong;XIONG Xun(Key Laboratory of Genetic Breeding and Reproduction of Plateau Mountain Animals of Ministry of Education/Key Laboratory of Animal Genetic Breeding and Reproduction of Guizhou Province,Guizhou University,Guiyang 550025,China;School of Animal Science,Guizhou University,Guiyang 550025,China;School of Life Sciences,Guizhou University,Guiyang 550025,China)

机构地区：[1]贵州大学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 [3]贵州大学生命科学学院,贵州贵阳550025

出　　处：《中国兽医学报》2023年第4期794-803,共10页Chinese Journal of Veterinary Science

基　　金：国家自然科学基金资助项目(31571279);教育部促美大资助项目([2015]2062号)。

摘　　要：旨在分析牛肌细胞增强因子MEF2C(myocyte enhancer factor 2C)基因的分子特征和在不同组织中的表达特性。以关岭牛为试验材料,扩增MEF2C基因CDS区,对MEF2C基因序列和蛋白结构进行分析,RT-qPCR检测MEF2C在不同组织中的表达特性。结果显示:牛MEF2C基因的CDS区全长为1326bp,编码441个氨基酸。MEF2C蛋白相对分子质量约为48kDa,理论等电点为7.71,主要以α-螺旋和无规则卷曲为主,含有1个MADS结构域,亚细胞定位显示MEF2C主要位于细胞核(60.90%)。牛MEF2C基因与牦牛同源性较近,与火鸡的同源性较远。RT-qPCR结果表明,MEF2C基因在关岭牛和杂交牛的7个组织中皆可表达,在肺脏和脾脏中差异极显著(P<0.01);心脏和肝脏以及背最长肌中差异显著(P<0.05);在成年牛的脾脏和心脏中表达量极显著高于犊牛(P<0.01);其他组织在犊牛阶段的表达量极显著高于成年牛阶段(P<0.01)。在成肌细胞分化过程中,MEF2C基因的表达量在0,12,24,48h呈逐渐上升趋势,不同时间段的表达量差异极显著(P<0.01)。结果表明MEF2C基因在关岭牛生长发育过程中发挥着重要的作用,为进一步探究MEF2C基因对关岭牛组织器官发育的影响,挖掘地方种质资源提供了数据支撑。The aim of this study was to analyze the molecular characteristics and expression characteristics of bovine muscle enhancer factor 2C(myocyte enhancer factor 2C,MEF2C)gene in different tissues.The CDS region of MEF2Cgene was amplified from Guanling cattle.The sequence and protein structure of MEF2Cgene were analyzed.The expression characteristics of MEF2Cin different tissues were detected by RT-qPCR.The results showed that the CDS region of bovine MEF2Cgene was 1326bp,encoding 441amino acids.The relative molecular weight of MEF2C protein was about 48kDa,and the theoretical isoelectric point was 7.71.It is mainly composed ofα-helix and random curl,and contained a MADS domain.Subcellular localization showed that MEF2C was mainly located in the nucleus(60.90%).Bovine MEF2Cgene has close homology with yak and far homology with turkey.The results of RT-qPCR showed that MEF2Cgene was expressed in all 7tissues of Guanling cattle and crossbred cattle,and the difference was very significant in lung and spleen(P<0.01).There were significant differences in heart and liver and longissimus dorsi muscle(P<0.05).The expression levels of MEF2Cin spleen and heart of adult cattle were significantly higher than those of calves(P<0.01).The expression levels of MEF2Cin other tissues in calf stage were significantly higher than that in adult stage(P<0.01).During the differentiation of myoblasts,the expression level of MEF2Cgene increased gradually at 0,12,24and 48h,and the expression level of MEF2Cgene differed significantly at different time periods(P<0.01).The results showed that MEF2Cgene played an important role in the growth and development of Guanling cattle,providing data support for further exploring the effects of MEF2Cgene on tissue and organ development of Guanling cattle and mining local germplasm resources.

关 键 词：关岭牛 MEF2C基因 基因表达 

分 类 号：S823.81[农业科学—畜牧学]