辽育白牛MSTN基因分子克隆及序列分析  

Liaoyu White Cattle MSTN Gene Molecular Clone And Sequence Analysis

作　　者：王娇娇[1] 张世伟 张丽君 王泽英[1] 罗光彬[1] 白文林[1] 

机构地区：[1]沈阳农业大学畜牧兽医学院,辽宁沈阳110886 [2]辽宁省畜牧业经济管理站,辽宁沈阳110032

出　　处：《现代畜牧兽医》2016年第10期21-27,共7页Modern Journal of Animal Husbandry and Veterinary Medicine

摘　　要：肌肉生长抑制素(简称MSTN)是肌肉发育负调控因子,为了实现将来对辽育白牛的分子育种,本试验对MSTN分子特征及其靶MicroRNA进行了分析。以辽育白牛为研究对象,利用分子生物学技术,设计特定引物对辽育白牛MSTN基因的编码区和调控区分段PCR扩增,并进行克隆和测序。借助生物信息学分析软件,寻找各个区域的单核苷酸多态性(SNP)并分析了MSTN基因分子特征、组织表达谱以及3'非翻译区靶标rniRNA。试验表明,MSTN基因5'调控区SNPs使启动子位置和转录因子结合情况发生变化,但编码区的突变未造成氨基酸改变。辽育白牛MSTN编码区序列全长为1 128 bp,编码375个氨基酸,该蛋白存在信号肽序列,为分泌性蛋白,疏水性较弱且不稳定。辽育白牛MSTN基因在不同组织中具有表达差异性,且肌肉中表达量最高。3'UTR存在肌肉生长发育功能miRNA,且比较保守。为开展辽育白牛分子育种研究奠定理论基础。Myostatin （MSTN） are negative regulatory factors of muscle development, in order to achieved the molecular breeding of liao white cattle in the future, so this study analysesed the MSTN molecular characteristics and target of micrornas. In this research, Liaoyu white cattle were studied by means of molecular biological technology. By designing the specific primers, Cod- ing regions and regulatory regions of MSTN gene in Liaoyu white cattle were PCR amplified, cloned and sequenced separately. Based on the results of bioinformatics analysis software, to find SNPs in all areas, to analyze molecular characteristics of MSTN gene, expression differences in differ- ent tissues and target miRNAs of 3＇ -UTR. Experimental results showed that the SNPs in 5p regula- tory region of MSTN gene could change the promoter position and the transcription factor binding conditions. But, the mutations in the Coding region had no effect on the protein composition. In Liaoyu white cattle, the full-length of Coding region sequences in MSTN gene was i128 bp, with coding 375 amino acids which was the weak hydrophobic and unstable secreted protein containing signal peptide sequence. In different tissues of Liaoyu white cattle, there were expression dif- ferences for MSTN gene, and highest expression was appeared in muscle tissue. Conservative target miRNAs existed in 3p -UTR of MSTN gene. Through this research, the theoretical foundation of mo- lecular breeding for Liaoyu white cattle can be laid.

关 键 词：辽育白牛 MSTN基因 MICRORNA 单核苷酸多态性 

分 类 号：S823.25[农业科学—畜牧学]