乐至黑山羊褪黑素合成酶AA-NAT基因cDNA克隆及序列分析  

Molecular cloning and sequence analysis of Lezhi Black-goat melatonin synthetase AA-NAT genes

作　　者：杨大前[1] 字向东[1] 王永[1] 杨春先[1] 邓一科[1] 卢建远[1] 付锡三 

机构地区：[1]西南民族大学青藏高原动物遗传资源保护与利用四川省重点实验室,四川成都610041 [2]乐至县科技局,四川乐至641500

出　　处：《西南民族大学学报（自然科学版）》2011年第4期575-581,共7页Journal of Southwest Minzu University(Natural Science Edition)

基　　金：国家科技支撑计划课题(2008BADC3B00)资助

摘　　要：研究旨在分析乐至黑山羊的AA-NAT基因与其它属种的差异.根据GenBank中绵羊AA-NAT基因序列(U29663.1)设计一对引物,以乐至黑山羊松果体总RNA为模板,通过RT-PCR技术对乐至黑山羊AA-NAT cDNA进行克隆测序和序列分析.研究表明:乐至黑山羊AA-NAT基因cDNA编码区全长为624bp,编码207个氨基酸.乐至黑山羊AA-NAT基因与绵羊的同源性最高(97.6%),其次是普通牛(95.51%)、人(83.33%)、黑猩猩(83.17%)、狗(81.41%)、猕猴(81.25%)和家鼠(80.77%).利用NJ法以该序列和推导的氨基酸序列构建的物种间分子系统进化树分析,结果表明乐至黑山羊先与绵羊聚为一类,再与普通牛聚为一类,而后与狗聚为一类,最后与鼠、人和猴聚为一类.The objective of this study is to identify the differences of Arylalkylamine N-Acetyltransferase （AA-NAT） gene between Lezhi Black-goat and other genus. The primers are designed according to the GenBank sequence of Ovis aries AA-NAT gene （U29663.1）. Total RNA is extracted from the pineal gland of the Lezhi Black-goat and the cDNA encoding AA-NAT is obtained by the reverse transcription PCR （RT-PCR）. The purified RT-PCR product is cloned into T vector, and then the sequence is analyzed. The results demonstrate that the 624bp product is the Lezhi Black-goat AA-NAT cDNA. The homologies of nucleotide sequences of the coding region of AA-NAT gene between the Ovis aries, Bos taurus, human, chimpanzee, dog, rhesus monkey, and house rat are 97.6, 95.51, 83.33, 83.17, 81.41, 81.25 and 80.77%. The molecular phylogenetic trees among species are constructed according to the nucleotide sequence and deduced amino acid sequence of the coding region of AA-NAT. The result indicates that Bos taurus, Lezhi Black-goat and Ovis aries assemble separately, and then assemble to a genus with dog, mouse, human and monkey.

关 键 词：乐至黑山羊 AA-NAT 分子克隆 序列分析 

分 类 号：S827[农业科学—畜牧学]