北京鸭Mx基因克隆及其在鸭呼肠孤病毒感染的鸭胚成纤维细胞中的表达分析

Cloning of Mx Gene of Beijing Duck and its Expression Analysis in DEF Incubated with Duck Reovirus

本试验旨在研究北京鸭Mx基因的结构及功能。利用干扰素诱导剂Poly（I∶C）诱导鸭胚成纤维细胞（DEF）,提取细胞总RNA,采用RT-PCR方法扩增北京鸭Mx基因全长编码区序列,并观察其在感染了鸭呼肠孤病毒（DRV）的DEF中的动态表达情况。北京鸭Mx基因序列分析结果显示,该基因编码区全长2 166bp,编码721个氨基酸残基。通过与GenBank中已登录的脊椎动物Mx基因进行核苷酸系统进化树分析,结果显示北京鸭Mx基因与野鸭Mx基因关系最近。北京鸭Mx序列与其他动物基因序列的比对结果显示,核苷酸同源性为47.8%~99.8%,氨基酸同源性为47.9%~99.4%。DRV孵育DEF后,Mx呈波动性表达。结果表明,本试验成功克隆了北京鸭Mx基因,预测分析证实其编码的蛋白具有脊椎动物Mx蛋白共有的结构特征,北京鸭Mx蛋白全基因的获得为下一步研究禽类Mx蛋白的抗病毒活性、作用机制及干扰素的监测奠定了基础。

The study was aimed to research the structure and function of Mxgene in Beijing duck.Full-length sequence of Beijing duck Mx gene was amplified by RT-PCR from the total RNA extracted from duck embryo fibroblast（DEF）induced by Poly（I∶C）.Furthermore,the expression of Mxgene in DEF infected with DRV was described.Sequence analysis indicated that the duck Mxgene contained an open reading frame（ORF）of 2 166 bp encoding aprotein of 721 amino acids.A phylogenetic tree based on Mx gene sequence was constructed.The results showed that Beijing duck Mxgene had the lowest distance with the gene from wild duck available in GenBank.Homology analysis showed that Beijing duck Mxgene nucleotides and deduced amino acids shared 47.8%to 99.8% and 47.9%to 99.4% homologies with those from other animals available in GenBank,respectively.Fluctuation expression of Beijing duck Mxgene was found with DEF incubated with duck reovirus.Duck Mxgene was successfully cloned and predicted with characteristic of typical structure of Mx family.Gaining Beijing duck Mxgene laid a foundation for further researching Mx protein antiviral activity,molecular mechanism and interferon monitoring of poultry.