鸭黄病毒囊膜E蛋白宿主细胞受体分子的筛选及功能鉴定

项目名称： 鸭黄病毒囊膜E蛋白宿主细胞受体分子的筛选及功能鉴定

项目编号： No.31201928

项目类型： 青年科学基金项目

立项/批准年度： 2013

项目学科： 畜牧学与草地科学、兽医学、水产学

项目作者： 李小康

作者单位： 河南科技大学

项目金额： 23万元

中文摘要： 鸭黄病毒是2010年4月份以来在我国出现的一种新型病毒，主要引起产蛋严重下降和脑炎，给养鸭业造成了重大损失。作为黄病毒属的新成员，鸭黄病毒感染宿主细胞的确切机制尚不清晰。本项目以鸭黄病毒囊膜E蛋白吸附宿主细胞为切入点，采用T7噬菌体展示技术筛选E蛋白宿主细胞受体分子，通过易感细胞病毒结合阻断试验、非易感细胞受体重建及病毒感染试验，对筛选的E蛋白受体分子进行鉴定。运用RNAi技术，抑制宿主细胞E蛋白受体分子的表达，并结合SILAC技术定量解析鸭黄病毒感染宿主细胞重要蛋白差异表达谱，分析重要差异表达蛋白与E蛋白受体的相关性，鉴定E蛋白受体分子在鸭黄病毒感染中的功能。本项目的研究，可从分子水平确证E蛋白受体的基因结构，阐明E蛋白受体分子在病毒感染宿主细胞过程中的作用，初步揭示鸭黄病毒囊膜E蛋白与宿主细胞的相互作用机制。对于深入研究鸭黄病毒的吸附侵入模式、组织嗜性及致病机理具有突破性的意义。

中文关键词： 鸭坦布苏病毒；囊膜蛋白；受体；抑制肽；噬菌体展示

英文摘要： Since April 2010, a severe outbreak of duck viral infection, with egg drop, feed uptake decline, ovary-viduct and encephalitis disease, has spread around the major duck-producing provinces of Fujian, zhejiang, jiangsu, henan, hebei, shandong, hubei in China. A new virus, called novel duck flavivirus(DFV), genomic sequencing of the isolated virus revealed that it is closely related to tembusu virus, have made huge economic losses for poultry industries. To date, there is no report overseas, some bionomics, classification, nucleotide sequencing and phylogenetic analysis were studied by the scholars in China, but DFV pathogenesis and the interaction mechanism between virus proteins and host proteins remain elusive. A causative strain named HN1 was isolated from sick shelducks by inoculating filtered spleen homogenates into the allantoic sacs of embryonated duck eggs. The virus was replicated well in embryonated duck and chicken eggs, and also in primary duck embryo fibroblasts . The major envelope glycoprotein(E) is known as the viral attachment protein(VAP) of flavivirus, it contains structural and functional elements that participate in the virus-host cell protein interaction and membrane fusion. In this study, E protein was cloned from HN1 strain and expressed to screening the putative receptor molecule(s) by Ph

英文关键词： duck tembusu virus；envelope(E) protein；receptor；inhibitory peptide；phage surface display