天祝白牦牛金属硫蛋白基因(MT-1)全序列分子克隆及生物信息学分析  

Cloning and Bioinformatic Analysis of Metallothionein-1 Gene(MT-1)in Tianzhu White Yak(Bosgrunneins)

作　　者：王雪莹[1] 姬菩忠 马琦[3] 

机构地区：[1]甘肃农业大学动物医学院,兰州730070 [2]兰州军区总医院,兰州730040 [3]甘肃农业大学生命科学技术学院,兰州730070

出　　处：《农业生物技术学报》2014年第7期883-891,共9页Journal of Agricultural Biotechnology

基　　金：甘肃省自然科学基金(No.1010RJZA154);甘肃省教育厅科研项目(No.0902-05)

摘　　要：金属硫蛋白-1基因(metallothionein-1,MT-1)是一类广泛存在于动物体内的金属结合蛋白,除维持金属动态平衡和重金属解毒外,还可参与清除自由基、拮抗电离辐射和抗应激等。为深入研究MT-1基因参与动物机体抗逆性的分子机制,本研究采用RT-PCR和RACE(rapid amplification of cDNA ends,RACE)技术获得天祝白牦牛(Bos grunneins)MT-1基因全长cDNA序列,运用生物信息学方法分析MT-1蛋白的理化性质、结构和不同物种间的同源性,并利用RT-qPCR方法分析MT-1基因在天祝白牦牛各个脏器和组织中的表达丰度。结果表明,天祝白牦牛MT-1基因cDNA序列全长为408 bp(GenBank登录号:KF770836),开放阅读框(ORF)长186 bp,编码61个氨基酸,氨基酸的分子量为5.981 kD;氨基酸序列分析显示,天祝白牦牛MT-1基因编码氨基酸序列与牛(Bos taurus)的同源性高达100%,同时与大多数哺乳动物相似;荧光定量PCR结果发现,MT-1 mRNA在天祝白牦牛8个不同脏器和组织中均有表达,心脏中表达量最高。天祝白牦牛MT-1基因的成功克隆为进一步研究该基因的功能提供了基础资料。Metallothionein-1（MT-1） is an ubiquitously dispersed metal-binding protein in animals,which serves in maintaining metal homeostasis and heavy metal detoxification,and also scavenges free radicals,resists ionize radiation and anti-stress.For further research on the molecular mechanism of MT-1 gene involved in stress resistance,full length cDNA sequence of MT-1 gene in Tianzhu white yak（Bos grunneins） was cloned by RT-PCR and RACE （rapid amplification of cDNA ends）.Physicochemical property,structure and homology of MT-1 encoded protein were analyzed by bioinformatic methods.Real-time PCR was used to analyze MT-1 mRNA expression profile in different tissues of Tianzhu white yak.The result showed that the full length of MT-1 cDNA was 408 bp （GenBank accession No.KF770836）,with an ORF （open reading frame） of 182 bp in length,which encoded 61 amino acids with a predicted molecular mass of 5.981 kD.The predicted amino acid sequences showed 100％ similarity with Bos taurus,and was highly homologous with most mammalians.The mRNA expression of MT-1 gene was found in 8 different tissues of Tianzhu white yak by Real-time PCR,the highest expression level was found in heart.Therefore,the full length cDNA of MT-1 gene was successfully obtained,which provides basic data for further study on its function.

关 键 词：金属硫蛋白1 天祝白牦牛 RACE 生物信息学分析 

分 类 号：S823.85[农业科学—畜牧学]