猪ADAR2基因全长cDNA克隆、序列特征及表达模式分析  

The Full-length cDNA Cloning,Sequence Characterization and Expression Pattern Analysis of Porcine ADAR2 Gene

作　　者：张跃博[1,2] 王立刚 侯欣华[1] 刘欣 颜华[1] 张龙超[1] 王立贤 ZHANG Yuebo;WANG Ligang;HOU Xinhua;LIU Xin;YAN Hua;ZHANG Longchao;WANG Lixian(Institute of Animal Science,Chinese Academy of Agricultural Sciences,Beijing 100193,China;College of Animal Science and Technology,Hunan Agricultural University,Changsha 410128,China)

机构地区：[1]中国农业科学院北京畜牧兽医研究所,北京100193 [2]湖南农业大学动物科学技术学院,长沙410128

出　　处：《畜牧兽医学报》2021年第3期620-629,共10页ACTA VETERINARIA ET ZOOTECHNICA SINICA

基　　金：中国农业科学院科技创新工程(ASTIP-IAS02);国家生猪产业技术体系(CARS-36);湖南省教育厅科学研究项目(19C0902)。

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ADAR 2基因cDNA全长6305 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。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 6305 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.

关 键 词：ADAR2 表达 CDNA克隆 RACE 猪 

分 类 号：S828.2[农业科学—畜牧学]