豫西脂尾羊CIDEa基因的克隆、序列分析与组织表达

doi:10.16431/j.cnki.1671-7236.2019.06.004

摘要/Abstract

摘要：

试验旨在对豫西脂尾羊诱导细胞凋亡DNA片段化因子45样效应因子A （cell death-inducing DNA fragmentation factor 45-like effectors A，CIDEa）基因进行克隆及生物信息学分析，并检测其在豫西脂尾羊不同组织中的表达。以豫西脂尾羊脂肪组织RNA为模板，采用RT-PCR方法扩增并克隆豫西脂尾羊CIDEa基因完整CDS区序列，对测序结果进行了相关生物信息学分析，并对CIDEa基因在豫西脂尾羊心脏、肝脏、肺脏、脾脏、肾脏、瘤胃、小肠、背最长肌、皮下和内脏脂肪组织中的表达进行了分析。结果显示，豫西脂尾羊CIDEa基因CDS区序列长660 bp，编码219个氨基酸；豫西脂尾羊与绵羊、水牛、黄牛、藏羚羊、猪和人的CIDEa基因同源性分别为99.1%、96.8%、96.2%、98.8%、85.0%和79.4%。蛋白理化性质分析表明，CIDEa蛋白分子质量为24.38 ku，理论等电点（pI）为9.12，属于碱性蛋白。跨膜结构和信号肽预测分析表明，CIDEa蛋白不含跨膜结构和信号肽。亚细胞定位分析表明，豫西脂尾羊CIDEa蛋白分布在细胞质（47.8%）、线粒体（26.1%）、细胞核（17.4%）、液泡（4.3%）和内质网（4.3%）。蛋白质三级结构预测发现，CIDEa蛋白结构具有2个α-螺旋、4个β-折叠及一些无规则卷曲。实时荧光定量PCR结果表明，CIDEa基因在豫西脂尾羊脂肪组织（皮下和内脏脂肪组织）中表达量较高，其他组织中表达量从高到低依次为小肠、肝脏、脾脏、瘤胃、心脏、肾脏、肺脏、背最长肌。这些结果可为进一步研究CIDEa基因在肉羊脂质代谢中的功能及羊肉品质调控提供基础资料。

关键词: 豫西脂尾羊; CIDEa基因; 克隆; 生物信息学分析; 组织表达

Abstract:

This study was aimed to clone cell death-inducing DNA fragmentation factor 45-like effector A (CIDEa),analyze the sequences by bioinformatics software,and detect the expression of CIDEa gene in different tissues of Yuxi fat-tailed sheep.Using RNA of the adipose tissue in Yuxi fat-tailed sheep as a template,the complete CDS sequence of CIDEa gene in Yuxi fat-tailed sheep was amplified and cloned,and the sequencing result was analyzed by bioinformatics.The expression of CIDEa gene in heart,liver,lung,spleen,kidney,rumen,small intestine,longissimus dorsal muscle,subcutaneous adipose and visceral adipose tissue of Yuxi fat-tailed sheep were analyzed.The results showed that the length of CDS sequence of CIDEa gene in Yuxi fat-tailed sheep was 660 bp and encoded 219 amino acids.The homology of CIDEa gene in Yuxi fat-tailed sheep were 99.1%,96.8%,96.2%,98.8%,85.0% and 79.4% with Ovis aries,Bubalus bubalis,Bos taurus,Pantholops hodgsonii,Sus scrofa and Homo sapiens,respectively.The physical and chemical properties of CIDEa protein showed that the molecular weight was 24.38 ku and the theoretical isoelectric point (pI) was 9.12,which belonged to alkaline protein.There was no transmembrane structure and signal peptide in CIDEa protein.The subcellular localization results showed that CIDEa protein was located in the cytoplasm (47.8%),mitochondria (26.1%),nuclear (17.4%),vacuolar (4.3%) and endoplasmic reticulum (4.3%).The tertiary structure of CIDEa protein mainly consisted of 2 alpha helices,4 beta foldings and random coils.Real-time quantitative PCR results showed that the expression of CIDEa gene in subcutaneous adipose and visceral adipose were obviously higher than other tissues,which followed by intestine,liver,spleen,rumen,heart,kidney,lung and longissimus dorsi muscle ranged from high to low.This results provided basic data for further study on the function of CIDEa gene in lipid metabolism and regulation of mutton quality in sheep.

Key words: Yuxi fat-tailed sheep; CIDEagene; cloning; bioinformatics analysis; tissue expression

中图分类号:

李君, 候霞飞, 梁文双, 邓红雨, 张桂枝, 刘太宇, 王笑笑, 权凯. 豫西脂尾羊CIDEa基因的克隆、序列分析与组织表达[J]. 《中国畜牧兽医》, 2019, 46(6): 1594-1602.

LI Jun, HOU Xiafei, LIANG Wenshuang, DENG Hongyu, ZHANG Guizhi, LIU Taiyu, WANG Xiaoxiao, QUAN Kai. Cloning,Sequence Analysis and Tissue Expression of CIDEa Gene in Yuxi Fat-tailed Sheep[J]. , 2019, 46(6): 1594-1602.

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