牦牛TBC1D7基因克隆、生物信息学及表达分析

Cloning,Bioinformatics Analysis and Expression Analysis of TBC1D7 Gene in Yak

为探究西藏牦牛Tre2-Bub2-CDC16结构域家族成员7(TBC1D7)基因的特征及结构,利用RT-PCR克隆了TBC1D7基因的CDS区序列,并对其进行了生物信息学分析,利用实时荧光定量PCR(qPCR)检测了TBC1D7基因在牦牛心脏、肝脏、脾脏、肺脏、肾脏、肌肉及脂肪组织的表达水平。结果表明,牦牛TBC1D7基因的CDS区全长为882 bp,共编码293个氨基酸;系统进化树分析结果表明,西藏牦牛与野牦牛的亲缘关系最近,与兔的最远;生物信息学分析结果表明,牦牛TBC1D7蛋白不存在信号肽和跨膜结构,属于亲水性蛋白,主要存在细胞核中,且该蛋白具有24个潜在的磷酸化位点,蛋白的高级结构由α-螺旋(65.53%)、延伸连(3.75%)、β-转角(3.75%)和无规则卷曲(29.96%)构成。qPCR检测结果显示,TBC1D7基因在西藏牦牛脾脏组织中的表达最高,极显著高于其他组织(P<0.01)。

In order to investigate the characteristics and structure of Tre2-Bub2-CDC16 structural domain family mem‐ber 7(TBC1D7)gene in Tibetan yak(Bos grunniens),the sequence of the CDS region of the TBC1D7 gene was cloned using RT-PCR.And bioinformatics analysis,real-time fluorescence quantitative PCR(qPCR)was used to detect the expression levels of TBC1D7 gene in heart,liver,spleen,lung,kidney,muscle and adipose tissues of yaks.The results showed that the CDS region of the yak TBC1D7 gene was 882 bp in length,encoded a total of 293 amino acids;The results of the phylogenetic tree indicated that the Tibetan yak was most closely related to the wild yak and more distantly related to the Oryctolagus cuniculus;The yak TBC1D7 protein was free of signal peptide and transmembrane structures,it was a hydrophilic protein and mainly found in the nucleus,it had 24 potential phosphorylation sites.The advanced structure of the protein was consisted ofα-helices(65.53%),stretching-links(3.75%),β-turns(3.75%),and irregular coiling(29.96%).qPCR results showed that the TBC1D7 gene was most highly expressed in the spleen tissues of Tibetan yaks,which was highly significantly higher than other tissues(P<0.01).