桑桑牦牛MYL7基因克隆及生物信息学分析

Cloning and Bioinformatics Analysis of MYL7 Gene in Sangsang Yak

克隆西藏桑桑牦牛肌球蛋白轻链7(Myosin light chain 7,MYL7)基因CDS区,进行生物信息学分析并检测该基因在各组织中的表达差异,为探究该基因功能提供一定的理论依据。以桑桑牦牛肌肉组织cDNA为模板,利用PCR技术克隆其CDS区序列,并利用多种在线生物软件及工具进行生物信息学分析。结果表明,桑桑牦牛MYL7基因CDS区长447 bp,共编码148个氨基酸,该基因编码的蛋白分子式为C743H1155N187O237S7,原子数为2329个,理论等电点为4.37,不稳定系数和总平均亲水性分别为36.02和-0.364,属于稳定的亲水性蛋白;具有特异性的磷酸化位点11个,其中5个丝氨酸、5个苏氨酸和1个酪氨酸。MYL7蛋白无信号肽和跨膜螺旋结构,属于非分泌性蛋白。通过亚细胞定位发现,桑桑牦牛MYL7基因在线粒体、细胞核、细胞质、细胞膜中均有所分布。蛋白高级结构主要是α-螺旋和无规则卷曲。通过构建系统进化树发现,桑桑牦牛与野牦牛亲缘关系最近,符合实际情况,说明MYL7基因在进化过程中具有一定的保守性。

The CDS region of Myosin light chain 7(MYL7)gene of Tibetan Sangsang yak was cloned,and bioinformatics analysis was carried out to detect the expression difference of this gene in various tissues,so as to provide a theoretical basis for exploring the function of this gene.The CDS region sequence was cloned by PCR using the cDNA of Sangsang yak muscle tissue as a template,and bioinformatics analysis was performed using a variety of online biological software and tools.The results showed that the CDS region of MYL7 gene was 447 bp in length and encoded 148 amino acids.The molecular formula of the protein encoded by this gene was C743H1155N187O237S7,the number of atoms was 2329,the theoretical isoelectric point was 4.37,the instability coefficient and the total average hydrophilicity were 36.02 and-0.364,respectively,which were stable hydrophilic proteins.There are 11 specific phosphorylation sites,including 5 serine,5 threonine and 1 tyrosine.MYL7 protein has no signal peptide and transmembrane helix structure,which belongs to non-secretory protein.Through subcellular localization,it was found that the MYL7 gene of Sangsang yak was distributed in mitochondria,nucleus,cytoplasm and cell membrane.The advanced structure of the protein was mainlyα-helix and random coil.By constructing a phylogenetic tree,it was found that Sangsang yak and wild yak had the closest relationship,which was in line with the actual situation,indicating that the MYL7 gene was relatively conservative in the evolutionary process.