山羊EDNRA基因编码区克隆与组织表达规律分析

doi:10.16036/j.issn.1000-2650.2019.06.015

摘要/Abstract

摘要： 【目的】旨在探究EDNRA基因在山羊心脏、肝、脾、肺、肾、大脑、网膜脂肪组织中的表达情况。【方法】采用克隆、测序的方法得到山羊EDNRA基因CDS全长的碱基序列,用生物信息学软件进行序列分析、亲水性/疏水性分析、潜在的信号肽位点分析、亚细胞定位分析、蛋白质的二级结构和三级结构预测及系统进化树分析,最后采用实时荧光定量PCR（qPCR）测定EDNRA基因在山羊心脏、肝、脾、肺、肾、大脑、网膜脂肪组织中mRNA的表达量。【结果】成功克隆了EDNRA的CDS序列,得到山羊EDNRA基因包含一个1 284 bp的最长开放阅读框。该序列编码427个氨基酸,含信号肽序列（第1到第20位氨基酸）以及7型跨膜受体同系物结构域。基因编码产物具有疏水性,属分泌蛋白,有35个磷酸化修饰位点。二级结构主要为α-螺旋。系统进化树分析表明,山羊EDNRA基因和绵羊、牛以及野牛的亲缘关系较近。qPCR结果表明,山羊EDNRA基因在心脏中的表达量最高,在肝、脾、肾、大脑、网膜组织中的表达量较低,而在肺中表达最少。【结论】EDNRA可能在心脏行使功能过程中发挥重要作用。

关键词: EDNRA基因, CDS区序列分析, 蛋白结构预测, 组织表达规律, 山羊

Abstract: 【Objective】 This study aim to investigate the expression of EDNRA gene in goat heart, liver, spleen, lung, kidney, brain and omental fat tissue. 【Method】 In this study, the full-length of the CDS region of goat EDNRA gene was cloned and sequenced. And bioinformatics softwares were used for sequence analysis, hydrophilic/hydrophobic analysis, potential signal peptide site analysis, subcellular localization analysis, protein secondary structure and tertiary structure prediction, and phylogenetic tree analysis. Finally, we used the real-time fluorescence Quantitative PCR (qPCR) to determine the mRNA expression of EDNRA gene in goat heart, liver, spleen, lung, kidney, brain and omental fat tissues. 【Result】 The CDS sequence of EDNRA was successfully cloned with the open reading frame as long as 1 284 bp. The CDS of EDNRA encodes 427 amino acids, including a signal peptide sequence (amino acids 1 to 20), and a type 7 transmembrane receptor homolog domain. The product of this gene was hydrophobic, which is part of the secretory protein, and had 35 phosphorylation modification sites. The α-helix was the main secondary structure. The results of phylogenetic tree analysis showed that the EDNRA gene of goat was closely related to sheep, cattle and bison. The results of qPCR showed that the goat EDNRA gene was highly expressed in the heart, but lowly expressed in the liver, spleen, kidney, brain and omental fat tissue, especially the least expression in the lung. 【Conclusion】 We speculated that EDNRA may play an important role in heart function.

Key words: EDNRA gene, CDS sequence analysis, protein structure prediction, patterns of tissue expr-ession, goat

中图分类号:

S811.5

王梦, 周靖宣, 占思远, 王林杰, 李利, 张红平, 仲涛. 山羊EDNRA基因编码区克隆与组织表达规律分析[J]. 四川农业大学学报, 2019, 37(06): 852-859.

WANG Meng, ZHOU Jingxuan, ZHAN Siyuan, WANG Linjie, LI Li, ZHANG Hongping, ZHONG Tao. The Cloning of Coding Region of EDNRA Gene and Its Expression Patterns in Goat[J]. Journal of Sichuan Agricultural University, 2019, 37(06): 852-859.

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