摘要
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（ Myl2） protein is an important member of the mammalian myosin.Sequencing of its gene,and analysis of its sequence features and expression profiles can help further understanding of its function. Using longissimus dorsi muscles of Small-tailed Han sheep as the experimental materials,the complete c DNA of ovine Myl2 gene was cloned using RT-PCR,5＇ RACE and3＇ RACE. The c DNA sequence was analyzed using related bioinformatics softwares. By using qRT-PCR and Western blotting,its mRNA and protein expressing levels were determined in various sheep tissues.We obtained 776 bp full-length c DNA and submitted to Gen Bank with the accession number of KJ710702. The c DNA contained 498 bp open reading frames（ ORF） and encoded protein composing 166 amino acid residues. The protein was predicted to have no signal peptide and disulfide bonds,but have several N-glycosylation and phosphorylation sites. More than half of the secondary structure of MYL2 was predicted to be α-helix. Sequence alignment showed that the sheep MYL2 protein shared more than 95%amino acid sequence similarity with those of Mus musculus,Homo sapiens,Rattus norvegicus,Sus scrofa,Bos Taurus,and so on. Both of the mRNA and protein expressing profiles indicated that Myl2 was expressed at highest abundance in the heart of sheep,followed by longissimus dorsi.
出处
《中国生物化学与分子生物学报》
CAS
CSCD
北大核心
2016年第12期1347-1353,共7页
Chinese Journal of Biochemistry and Molecular Biology
基金
潍坊学院博士基金(No.2015BS04)
山东省羊产业创新团队(No.SDAIT-0901101)
山东省自然科学基金(No.ZR2016CB28)资助~~
关键词
绵羊
肌球蛋白轻链2
克隆
特征
表达谱
sheep
myosin light chain 2
clone
molecular characterization
expressing profile
