不同羊毛纤维直径细毛羊MYL6基因克隆及差异表达研究  

Study on Clone and Differential Expression of MYL6 Gene in Fine-Wool Sheep with Different Fiber Diameter

作　　者：付雪峰[1] 黄锡霞[2] 刘春洁[1,2] 徐新明[1] 张艳花[1] 吴伟伟[1] 于丽娟[1] 石刚[1,3] 田可川[1] 

机构地区：[1]新疆畜牧科学院畜牧研究所,新疆绒毛用羊遗传育种与繁殖实验室,新疆乌鲁木齐830011 [2]新疆农业大学动物科学学院,新疆乌鲁木齐830052 [3]新疆科创畜牧繁育中心,新疆乌鲁木齐830000

出　　处：《中国畜牧杂志》2016年第13期5-11,共7页Chinese Journal of Animal Science

基　　金：田可川自然科学基金资助项目(31460593);黄锡霞自然科学基金资助项目(31360543);国家绒毛用羊产业技术体系建设专项(CARS-40);农业科研杰出人才及绒毛用羊研究创新团队;新疆绒毛用羊遗传育种与繁殖重点实验室专项(XJYS1105);家养动物种资质源平台:自治区公益性科研院所基本科研业务经费项目

摘　　要：试验以苏博美利奴羊为研究对象,探讨肌球蛋白轻链6(MYL6)基因在不同羊毛纤维直径的苏博美利奴羊中的表达模式。通过克隆肌球蛋白轻链6(MYL6)基因,结合生物信息学方法对其进行遗传进化和理化性质分析,且预测其二级蛋白结构,又通过RT-PCR相对定量方法以2^(-△△ct)值检测MYL6基因在苏博美利奴羊上的表达规律。结果表明:苏博美利奴羊MYL6基因大小为152036915 bp,MYL6基因的CDS全长序列为696 bp,共编码了105个氨基酸,与绵羊、山羊、牛、猪、老鼠、兔子、鸡、狗和斑马鱼的CDS同源分别为96%、95%、40%、40%、40%、40%、0%、60%、0%。分别在1、220~336、415~864 bp位点上有3个开放式阅读框(ORF)。遗传理化性质研究显示MYL6蛋白含有1个信号肽,在苏氨酸上有5个磷酸化位点、6个N-糖基化位点和3个O-糖基化位点,可能是一种非分泌性蛋白;MYL6蛋白的二级结构以无规则卷曲为主,β-折叠区域占-2.00%;扭曲度占-0.16%;等电荷在-1.60~0.05。研究结果表明,苏博美利奴羊MYL6基因在超细型细毛羊皮肤组织表达量显著高于细型细毛羊的表达量(P<0.01,差异倍数为2.048)。本研究将为开展肌球蛋白轻链基因在细毛羊皮肤转录水平上的研究提供数据理论基础。The purpose of this study was to investigate the expression patterns of myosin light chian gene（6） （MYL6） in Subo Merino of different wool fiber diameter. The MYL6 was cloned by electronic means. And then, the genetic evolution and physicochemical property of MYL6 were analyzed by bioinformatic methods. Meanwhile, the secondary protein structure of MYL6 was predicted, and MYL6 gene expression distribution were detected by relative quantitative RT-PCR method with 2-△△ct value in different individuals of Subo Merino. The results showed that the MYL6 gene size was 152036915bp in Subo Merino. The CDS full-length sequence of MYL6 gene is 696bp, encoding 105 amino acids, and the CDS isogeny with sheep, goats, cows, pigs, mice, rabbits, chickens, dogs and zebrafish homologous were 96%, 95%, 40%, 40%, 40%, 40%, 0%, 60%, 0%, and have three open reading frame （ORF） on lbp, 220-336bp, 415-864bp loci, respectively. Phylogenetic analysis indicated that the MYL6 protein transmembrane domain have 1 signal prediction, 5 phosphorylation sites, 6 N-glycosylation sites, 30-glycosylation sites in threonine, and it may be not a kind of secretory proteins. The MYL6 protein secondary structure is given priority to with random curl,β-sheet is accounted 2.00%, torsion resistance is -0.16%, equal charge between -1. 60 in 0.05. The MYL6 gene expression levels in superfine type skin tissue were significantly higher than fine type Subo Merino Sheep （P〈 0.01, 2.048 fold difference ）. This results will provide some data and theory basis for the Myosin light chain gene research at transcription level in fine wool sheep.

关 键 词：苏博美利奴羊 MYL6基因 生物信息学 RT-PCR 转录水平 

分 类 号：S827.2[农业科学—畜牧学]