延边黄牛DGATs基因克隆、生物信息学及组织表达分析

doi:10.16431/j.cnki.1671-7236.2021.09.006

摘要/Abstract

摘要： 试验旨在克隆延边黄牛二酯酰甘油酰基转移酶（diacylglycerol acyltransferase，DGATs）两种亚型（DGAT1和DGAT2）的CDS区核苷酸序列，并根据生物信息学对两种亚型的氨基酸序列进行分析，以及探讨其在延边黄牛各组织中的表达规律。采用RT-PCR和实时荧光定量PCR技术分别进行DGATs基因CDS区的扩增、克隆以及其在延边黄牛8个组织中mRNA表达量的检测，利用NCBI中BLAST将其与其他物种进行相似性分析，并构建系统进化树；通过在线工具对其编码蛋白的理化性质、一级结构及高级结构进行预测。结果显示，DGAT1和DGAT2基因CDS区序列长度分别为1 470和1 086 bp，分别编码489和361个氨基酸；二者均为稳定的疏水性蛋白。DGAT1存在25个潜在的磷酸化位点、1个N-糖基化位点和8个跨膜结构域；DGAT2存在28个潜在的磷酸化位点、2个N-糖基化位点和1个跨膜结构域。二者均不存在信号肽，即不属于分泌蛋白。DGAT1主要是通过α-螺旋和无规则卷曲连接，而DGAT2以α-螺旋、无规则卷曲和延伸链连接为主。延边黄牛DGAT1和DGAT2基因分别在延边黄牛小肠和脂肪组织中表达量最高，且显著高于其他组织（P<0.05）。本试验结果对进一步研究延边黄牛DGATs基因以及探究其在脂肪沉积过程中的作用机制具有重要意义。

关键词: 延边黄牛; DGATs基因; 克隆; 生物信息学; 表达

Abstract: The purpose of this study was to clone the CDS nucleotide sequences of two subtypes (DGAT1 and DGAT2) of diacylglycerol acyltransferase (DGATs) in Yanbian Yellow cattle, analyze the amino acid sequences of two subtypes according to bioinformatics, and explore the expression patterns of two subtypes in Yanbian Yellow cattle. RT-PCR and Real-time quantitative PCR were used to amplify and clone DGATs gene CDS and detect its mRNA expression in 8 tissues of Yanbian Yellow cattle, respectively. BLAST in NCBI was used to analyze its similarity with other species and construct phylogenetic tree. A series of online tools were used to predict the physicochemical properties, primary structure and higher structure of the proteins encoded by it. The results showed that the CDS of DGAT1 and DGAT2 genes were 1 470 and 1 086 bp, coding 489 and 361 amino acids, respectively. Both of them were stable hydrophobic proteins. There were 25 potential phosphorylation sites, 1 N-terminal glycosylation site and 8 transmembrane domains for DGAT1, and 28 potential phosphorylation sites, 2 N-terminal glycosylation sites and 1 transmembrane domain for DGAT2. There was no signal peptide in both of them, so they were not secretory proteins. DGAT1 was dominated by alpha helix and random coil, while DGAT2 was dominated by alpha helix, random coil and extended chain. The expression levels of DGAT1 and DGAT2 genes in small intestine and adipose tissue of Yanbian Yellow cattle were the highest, which were significantly higher than other tissues (P<0.05). The results of this study were of great significance to further study the DGATs gene of Yanbian Yellow cattle and explore its mechanism in the process of fat deposition.

Key words: Yanbian Yellow cattle; DGATs gene; cloning; bioinformatics; expression

中图分类号:

郭盼盼, 金鑫, 孙建富, 李强, 李香子, 严昌国. 延边黄牛DGATs基因克隆、生物信息学及组织表达分析[J]. 中国畜牧兽医, 2021, 48(9): 3158-3170.

GUO Panpan, JIN Xin, SUN Jianfu, LI Qiang, LI Xiangzi, YAN Changguo. Cloning,Bioinformatics and Tissue Expression Analysis of DGATs Genes in Yanbian Yellow Cattle[J]. China Animal Husbandry and Veterinary Medicine, 2021, 48(9): 3158-3170.

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