猪ADAR1基因cDNA全长克隆、序列信息及组织表达分析

doi:10.11843/j.issn.0366-6964.2019.06.003

摘要/Abstract

摘要： 旨在克隆猪作用于RNA的腺苷脱氨酶1基因（ADAR1） cDNA全长序列，检测该基因在大白猪不同组织及不同日龄背膘中的表达情况。本研究利用RACE（rapid-amplification of cDNA ends）克隆大白猪ADAR1基因mRNA全长序列，并对其进行生物信息学分析；采用荧光定量PCR方法检测ADAR1在不同组织中的表达水平，并探究不同日龄背膘中ADAR1的表达规律。结果表明，猪ADAR1基因cDNA全长6 259 bp，编码1 145个氨基酸，与人、黑猩猩、猕猴、长臂猿、黄牛、山羊和绵羊的氨基酸序列一致性均不低于85%。该基因编码的蛋白具有2个Zα结合结构域，3个双链RNA结合结构域和1个脱氨酶结构域。ADAR1在心、肝、脾、肺、肾、脑、肌肉、小肠和背部脂肪组织以及7、60、120和180日龄个体背膘组织中均表达，其表达量总体上表现出随个体发育先下降后上升的趋势。综上所述，本研究成功克隆了猪ADAR1基因cDNA全长序列，发现其在猪体内广泛表达，并且在不同日龄猪背膘组织中表达水平存在差异，为今后深入研究ADAR1的功能奠定了理论基础。

关键词: ADAR1, 表达, 基因克隆, RACE, 猪

Abstract: This experiment was conducted to clone the full-length cDNA of porcine ADAR1 gene, and explore its expression in different tissues and backfat of pigs with different ages.The cDNA full-length sequence of ADAR1 gene was cloned using rapid-amplification of cDNA ends (RACE) and analyzed by bioinformatics. The real-time PCR was used to detect ADAR1 mRNA expression in different tissues and backfat of Large White pigs with different ages. A 6 259 bp cDNA sequence of porcine ADAR1 was cloned, which encoded 1 145 amino acids. Its amino acid sequence shared an identity equal or more than 85% with other mammals including human, chimpanzee, macaque, gibbon, cow, goat and sheep. The predicted ADAR1 had 2 Zα binding domains, 3 double-stranded RNA binding domains and a deaminase domain. The real-time PCR result showed that ADAR1 was expressed in heart, liver, spleen, lung, kidney, brain, muscle, small intestine and backfat. Furthermore, it was expressed in backfat of individuals with the ages of 7, 60, 120 and 180 days, showing an overall trend of going down firstly and going up then with the individual development. In this study, the full-length cDNA sequence of ADAR1 gene was successfully cloned in pigs, which was widely expressed in pigs and its expression levels were different in backfat of pigs with different ages. These findings provided a theoretical foundation for further function study of ADAR1.

Key words: ADAR1, expression, gene clone, RACE, swine

中图分类号:

S828.2

张跃博, 欧阳峰正, 王立刚, 侯欣华, 刘欣, 颜华, 张龙超, 王立贤. 猪ADAR1基因cDNA全长克隆、序列信息及组织表达分析[J]. 畜牧兽医学报, 2019, 50(6): 1135-1144.

ZHANG Yuebo, OUYANG Fengzheng, WANG Ligang, HOU Xinhua, LIU Xin, YAN Hua, ZHANG Longchao, WANG Lixian. The Full-length Cloning, Sequence Information and Expression Analysis of Porcine ADAR1 Gene[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2019, 50(6): 1135-1144.

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