从规模化养鸭场分离到4株鸭坦布苏病毒,成功克隆了包膜蛋白(E蛋白)基因片段,并对其进行了序列测定、分析和表达。结果表明,E基因全长1503bp,编码501个氨基酸,序列高度保守。进化分析显示,鸭坦布苏病毒属蚊媒黄病毒类恩塔亚病毒群,与其他坦布苏病毒及近期发现的BYD病毒关系最近。采用原核表达系统对E基因进行了表达,蛋白大小54300,主要以包涵体形式存在。随后,对E蛋白外功能区序列和3D结构进行了分析,在已解析结构的黄病毒中与wNVE结构最为相似。研究结果为该病的防控和基因工程疫苗的研制奠定了基础。
Envelop protein (E protein) genes of four duck Tembusu virus (TMUV) isolated from duck farm were cloned, sequenced and expressed. E gene comprised 1 503 nucleotides, encoding 501 amino acid. Phylogenetic analysis shows that duck TMUV belongs to mosquito-born flavivir- us,NTAV group, similar to other Tembusu and BYD virus. The molecular weight of E protein was 54 300,which was expressed in pET-21a/BL21(DE3) system. The sequence and three-dimensional structure of ectodomain of duck TMUV were most similar to WNV E protein. The result was useful for the control and the development of molecular vaccine of duck TMUV.