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牦牛PID1基因克隆、生物信息学及组织表达分析
Cloning, Bioinformatics and Tissue Expression Analysis of PID1 Gene in Yak
【摘要】 为研究牦牛磷酸酪氨酸互作结构域1基因(PID1)的结构及功能,并探究其在各组织中的表达情况,以桑桑牦牛脂肪组织cDNA为模板克隆桑桑牦牛PID1基因CDS区序列,并对其进行生物信息学分析。同时,采用实时荧光定量PCR方法(RT-qPCR),对牦牛的7个不同组织包括心、肝、脾、肺、肾、背部肌肉及脂肪进行PID1基因表达水平的定量。结果表明,牦牛PID1基因的编码区长度是654 bp,编码217个氨基酸;通过同源性比对,结果发现,牦牛与野牦牛之间的亲缘关系最为接近,相似度达到100%;经过蛋白质分析发现,牦牛PID1蛋白的分子质量约为24.84 ku,理论等电点为6.30。根据不稳定系数计算结果显示,其不稳定性较高(47.96),属于一种不稳定蛋白质。该蛋白内含有1个N-糖基化位点和23个磷酸化位点,无信号肽和跨膜结构。RT-qPCR试验结果表明,在各个组织中都能检测到PID1基因的表达情况,其中在肺部表达量最高。该研究克隆了牦牛PID1基因,并对其蛋白结构进行了分析,同时还研究了该基因在牦牛组织中的表达情况。为进一步探究PID1基因在牦牛脂肪沉积过程中的作用提供了初步数据。
【Abstract】 To study the structure and function of phosphotyrosine interaction domain 1(PID1) gene in yak, and to explore its expression in various tissues.The CDS region of Sangsang yak PID1 gene was cloned using Sangsang yak adipose tissue cDNA as template, and the sequence was analyzed bioinformatically.Meanwhile, the relative expression level of PID1 gene was detected in seven tissues of yak namely heart, liver, spleen, lung, kidney, longissimus dorsi muscle and adipose tissue by Real-time Fluorescence Quantitative PCR(RT-qPCR).The results showed that the PID1 gene in yaks had a coding region length of 654 bp, which encoded 217 amino acids.Homology comparison showed that yak and wild yak were closely related, and the similarity reached 100%.The molecular weight of yak PID1 protein was about 24.84 ku and the theoretical isoelectric point was 6.30.According to the calculation results of instability coefficient, the instability of the protein was high(47.96),and it belonged to an unstable protein.The protein had one N-glycosylation site and 23 phosphorylation sites, with no signal peptide or transmembrane structure.The results of RT-qPCR showed that the expression of PID1 gene could be detected in all tissues, with the highest expression in the lung.The yak PID1 gene was cloned and its protein structure was analyzed.The expression of PID1 gene in yak tissue was also studied.Further study on the role of PID1 gene in yak fat deposition provided preliminary data.
- 【文献出处】 华北农学报 ,Acta Agriculturae Boreali-Sinica , 编辑部邮箱 ,2024年06期
- 【分类号】S823.85
- 【下载频次】76