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肌球蛋白轻链2蛋白是哺乳动物肌球蛋白的重要成员之一。获得其基因序列,并对其特征和表达进行分析,可为进一步研究功能奠定基础。本研究以小尾寒羊背最长肌为试验材料,采用RACE等方法对绵羊肌球蛋白轻链2基因的c DNA序列进行克隆和测序、利用相关生物学软件对所得c DNA序列进行生物信息学预测、并利用qRT-PCR和Western印迹法对其在绵羊各种组织中的表达进行分析。结果获得该基因c DNA序列全长为776 bp,提交至Gen Bank中获得相应的登录号为KJ710702;该序列中的498 bp的开放读码框编码含有166个氨基酸残基的蛋白质。预测发现该蛋白质无信号肽和二硫键,但存在N-糖基化和磷酸化位点;二级结构中以α-螺旋为主;蛋白质序列比较发现绵羊MYL2与小鼠、人、大鼠、猪、牛等哺乳动物的同源性均在95%以上。mRNA和蛋白质表达谱均显示该基因在绵羊心肌中表达量最高,其次为背最长肌。
Myosin light chain 2 protein is one of the important members of mammalian myosin. Obtaining its gene sequence and analyzing its characteristics and expression can lay a foundation for further study on the function. In this study, the tail muscle of Little Tail Han sheep was used as experimental material, and the cDNA sequence of sheep myosin light chain 2 gene was cloned and sequenced by RACE method. The bioinformatics Predicted and analyzed for their expression in various tissues of sheep using qRT-PCR and Western blotting. Results The full-length cDNA sequence of this gene was 776 bp and was submitted to Gen Bank to obtain the corresponding accession number KJ710702. The 498 bp open reading frame in this sequence encodes a protein with 166 amino acid residues. It was predicted that there was no signal peptide and disulfide bond in this protein, but there were N-glycosylation and phosphorylation sites. The secondary structure consisted mainly of α-helix. The protein sequence comparison showed that MYL2 and mouse, human and rat , Pigs, cattle and other mammals are more than 95% homology. The mRNA and protein expression profiles showed that the gene was most expressed in sheep myocardium, followed by the dorsal longissimus muscle.