三黄鸡NHE1基因的克隆和序列分析  

Cloning and sequencing of NHE1 genes in Chinese Yellow Chicken

作　　者：黄志强[1] 安玉甫[1,2] 于梦[1] 童双[1] 刘荣昌[1] 王衡[1] 宁章勇[1] 

机构地区：[1]华南农业大学兽医学院病理教研室,广东广州510642 [2]滁州温氏畜牧有限公司,安徽滁州239500

出　　处：《畜牧与兽医》2013年第11期35-38,共4页Animal Husbandry & Veterinary Medicine

基　　金：广东省自然科学基金(7300744)

摘　　要：钠氢离子交换泵1(NHE1)是一种跨膜蛋白,作为已经被证明的J亚群白血病病毒受体在该病的进程中起着关键性的作用。为全面了解NHE1的基因及其编码蛋白的生物学信息,对三黄鸡NHE1基因进行了克隆和序列分析。参考原鸡NHE1基因序列设计引物,采用PCR技术分段克隆三黄鸡NHE1基因序列,并对其进行序列分析。结果表明,克隆得到的三黄鸡NHE1基因全长2 412 bp,含有1个2 412 bp的完整开放式阅读框,编码803个氨基酸。其CDS编码区的核苷酸序列与原鸡、鹌鹑、人、猪、鼠、牛、弓形虫的NHE1基因对应序列的同源性分别为99.8%、92%、78.1%、73.5%、80%、81.8%、40.9%,推导的氨基酸序列同源性分别为99.5%、93.2%、79.3%、79.6%、78.9%、78.7%、26.8%。进化树分析发现,物种内的核苷酸序列的同源性很高,种间同源性较低。生物信息学预测NHE1蛋白结构发现,NHE1蛋白多肽链N端含有12个跨膜结构,有37个磷酸化位点和1个特异性蛋白激酶磷酸化位点。The Na^＋/H^＋ exchanger 1 （ NHE1 ）, a transmembrane protein, has been proved to play a critical role in the progression of the pathogenesis of subtype J avian leukosis as the receptor of ALJ-V. For a comprehensive understanding of the biological information of chick-en NHE1 gene and its encoding protein, the current study was designed to analyse sequence of NHE1 Gene in Chinese yellow chicken. Based on the sequence of the NHE1 of the chicken reported in NCBI, 2 pairs of primers were designed and the primer were used to amplify the Chi- nese yellow chicken NHE1 gene by PCR. The bioinformatics analysis of NHE1 was summarized. The results showed that full length of the gene of NHE1 was 2412 bp, including the 2412 bp of open reading frame and encoded 803 amino acid residues deduced protein. This nucle- otide sequence of CDS shared 99. 8%, 92%, 78.1%, 73.5%, 80%, 81.8% and 40. 9% homology with the NHE1 mRNA of Gallus gal-lus, Coturnix coturnix, Homo sapiens, Susscrofa, Musmusculus, BosTaurus and Toxoplasma gondii, respectively and the homology of de-duced amino acids were 99.5%, 93.2%, 79.3%, 79. 6%, 78.9%, 78.7% and 26. 8%, respectively. The biologic softwares were used to predict the bioinformatics of NHE1, which showed that the NHE1 was a kind of transmembrance protein and have 12 transmembrane do-mains in chicken. There were 37 phosphorylattion sites and one phosphorylation site of specific protein kinase in NHE1 protein.

关 键 词：NHE1 鸡 克隆 J亚群白血病受体 序列分析 

分 类 号：Q811.4[生物学—生物工程]