牦牛NRAMP1基因3′UTR克隆及其miRNA生物信息学分析

Cloning and miRNA bioinformatics analysis of yak NRAMP1 gene 3′UTR

为了解牦牛天然抗性相关巨噬细胞蛋白1(natural resistance associated macrophage protein 1,NRAMP1)基因3′UTR序列多态性及可能具有靶向关系的miRNAs,试验选取5个地区总计325头牦牛的血液样本,构建5个DNA混合池,对牦牛NRAMP1基因3′UTR进行PCR扩增,利用反向测序方法对扩增产物进行测序,使用DNAMAN 8.0软件将扩增序列与野牦牛、瘤牛、家牛和水牛NRAMP1基因3′UTR序列进行对比分析,并运用TargetScan和miRWalk数据库对靶向牦牛NRAMP1基因3′UTR的miRNAs进行预测。结果表明:成功扩增出牦牛NRAMP1基因3′UTR,扩增片段含有多个短串联重复序列,并检测出3个突变碱基;其与野牦牛、瘤牛/家牛和水牛的NRAMP1基因3′UTR同源性分别为100%、99%、99%、98%;牦牛NRAMP1基因3′UTR的可能靶向miRNAs基因共有11个。

In order to understand the polymorphism of 3′UTR region of natural resistance associated macrophage protein 1(NRAMP1) gene in yak and miRNAs that may be targeting to NRAMP1 gene, the 3′UTR region of NRAMP1 gene in 325 yaks from 5 regions was amplified by PCR using five DNA mixing pools.The amplified products were sequenced by reverse sequencing and DNAMAN 8.0 software was used to compare and analyze the 3′UTR region of NRAMP1 gene in wild yak, zebu cattle, domestic cattle and buffalo cattle, and the miRNAs targeting the 3′UTR region of NRAMP1 gene in yak were predicted by using TargetScan and miRWalk prediction database.The results showed that the 3′UTR region of NRAMP1 gene in yak contained multiple short tandem repeats, and three base mutations were detected. The homology of 3′UTR region of NRAMP1 gene with wild yak, zebu cattle, domestic cattle and buffalo cattle was 100%, 99%, 99% and 98%.There are 11 possible miRNAs predicted to target genes in the 3′UTR region of yak NRAMP1 gene.