猪ADAR2基因全长cDNA克隆、序列特征及表达模式分析

doi:10.11843/j.issn.0366-6964.2021.03.006

摘要/Abstract

摘要： 旨在克隆猪作用于RNA的腺苷脱氨酶2基因（ADAR2）全长cDNA序列，同时对该基因在猪不同组织中的表达规律进行探索。利用RACE （rapid-amplification of cDNA ends）对大白猪ADAR2基因mRNA全长序列进行克隆，并进行生物信息学分析；用荧光定量PCR方法检测35日龄大白猪心、肝、肺、肾、脾、脑、小肠、背最长肌和背部脂肪9种组织中ADAR2的表达水平。结果表明，猪ADAR2基因cDNA全长6 305 bp，共包含12个外显子，编码704个氨基酸，与人、黑猩猩、猕猴、长臂猿、黄牛、山羊和绵羊的CDS区核酸序列和氨基酸序列的一致性均在84%以上。该基因编码的蛋白含有2个双链RNA结合基序和一个脱氨酶结构域。猪ADAR2在检测的各组织中均表达，其中在肺中的表达量最高。综上所述，本研究成功克隆了猪ADAR2基因全长cDNA序列，并且发现其在猪体内广泛表达，为深入研究ADAR2的功能奠定了良好的基础。

关键词: ADAR2, 表达, cDNA克隆, RACE, 猪

Abstract: This study aimed to clone the full-length cDNA of the porcine ADAR2 gene and explore its expression pattern in different tissues of pigs. Rapid-amplification of cDNA ends (RACE) was used to clone the full-length cDNA sequence of the ADAR2 gene in Large White pigs and the sequence was analyzed by bioinformatics. Real-time PCR was used to detect the ADAR2 mRNA expression in the heart, liver, lung, kidney, spleen, brain, small intestine, muscle and backfat of 35-day-old pigs. Porcine ADAR2 cDNA sequence of 6 305 bp was cloned, which contained 12 exons and encoded 704 amino acids. The nucleic acid and amino acid sequences shared a high identity (>84%) with that of other mammals including human, chimpanzee, macaque, gibbon, cow, goat and sheep. The deduced ADAR2 had two double-stranded RNA binding motifs and an adenosine deaminase domain. Real-time PCR results showed that the ADAR2 expressed in all the detected tissues, and had the highest expression in the lung. The full-length cDNA sequence of porcine ADAR2 gene was successfully cloned, and it was widely expressed in tissues of pigs. These findings provide a solid foundation for further function study of ADAR2.

Key words: ADAR2, expression, cDNA clone, RACE, swine

中图分类号:

S828.2

张跃博, 王立刚, 侯欣华, 刘欣, 颜华, 张龙超, 王立贤. 猪ADAR2基因全长cDNA克隆、序列特征及表达模式分析[J]. 畜牧兽医学报, 2021, 52(3): 620-629.

ZHANG Yuebo, WANG Ligang, HOU Xinhua, LIU Xin, YAN Hua, ZHANG Longchao, WANG Lixian. The Full-length cDNA Cloning, Sequence Characterization and Expression Pattern Analysis of Porcine ADAR2 Gene[J]. Acta Veterinaria et Zootechnica Sinica, 2021, 52(3): 620-629.

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