牦牛GOLT1B基因克隆及生物信息学分析

Cloning and Bioinformatics Analysis of GOLT1B Gene in Yak

为研究牦牛GOLT1B基因的结构及功能,以美仁牦牛脂肪组织cDNA为模板,克隆牦牛GOLT1B基因的编码区序列(Codingsequence,CDS),并且利用该基因序列进行生物信息学分析、结构域预测及蛋白理化性质预测。结果表明,美仁牦牛GOLT1B基因的CDS区全长417 bp,共编码138个氨基酸;牦牛GOLT1B结构域预测显示,在GOLT1B氨基酸序列的第11~114位有1个Pfam结构域的Got1家族;GOLT1B蛋白预测结果表明,跨膜结构域有3处,分别位于氨基酸序列的第13~30位、第34~56位以及第69~91位,无信号肽区域,其氨基酸分子质量15425.69 Da,原子总数2232个,等电点10.36,不稳定系数25.43,为稳定的蛋白质,脂肪系数117.10,总平均亲水系数1.087,为疏水蛋白,共有10个磷酸化位点;同时,亚细胞定位结果表明,美仁牦牛GOLT1B基因主要分布于细胞外基质和细胞壁中;系统进化树表明:美仁牦牛与野牦牛和欧洲牛的亲缘关系最近。

In order to study the structure and function of GOLT1B gene in yak,the coding region sequence(CDS)of GOLT1B gene in yak was cloned using the cDNA of fat tissue of Meiren yak as the template,and the bioinformatics analysis and protein physicochemical properties prediction were carried out using the gene sequence.The results showed that the total length of the CDS region of the GOLT1B gene in Meiren yak was 417 bp,encoding a total of 138 amino acids;there was a Got1 family with a Pfam domain at positions 11-114 of the GOLT1B amino acid sequence;there were three transmembrane domain,which were located at positions 13-30,34-56,and 69-91 of the amino acid sequence,without signal peptide region.Its amino acid molecular weight was 15425.69 Da,the total number of atoms was 2232,the isoelectric point was 10.36,and the instability coefficient was 25.43,it was a stable protein,the fat coefficient was 117.10,and the total average hydrophilicity coefficient was 1.087,which was hydrophobic protein,with a total of 10 phosphorylation sites;the GOLT1B gene of Meiren yak was mainly distributed in the extracellular matrix and cell wall.Meiren yak had the closest relationship with wild yak and European cattle.