【作者】 李丽；

【导师】 常城；

【作者基本信息】 兰州大学 ， 动物学， 2008， 硕士

【摘要】 禽流感病毒非结构蛋白1(NS1蛋白)全长约为230个氨基酸,理论分子量约为26KDa,主要包括两个功能结构域,即N-末端的RNA结合结构域和和C-末端的效应结构域。NS1蛋白是一个多功能的病毒蛋白,它不仅影响着该病毒其他基因的表达,更能通过与宿主多种细胞因子的相互作用干预宿主细胞的正常功能,抵抗宿主的抗病毒系统。NS1蛋白因此被认为是禽流感病毒的一个重要毒力因子。为了寻找与NS1相互作用的其它宿主蛋白以及试图探索NS1的新功能,最终阐明禽流感的致病性机制,我们利用酵母双杂交系统,以NS1为诱饵蛋白,从人胸腺cDNA文库中筛到了8个宿主因子可与NS1蛋白相互作用,我们选择U蛋白作为我们研究的目的蛋白,并作了点对点酵母双杂交回转实验。结果发现两者确实是互作的,并且互作的部位位于NS1蛋白的RNA结合结构域上。同时我们利用GST Pull-down进一步验证了二者的相互作用。在研究当中还发现,高致病性禽流感病毒H5N1亚型NS1蛋白的80-84位氨基酸缺失可能对禽流感的致病性很重要,这提示针对H5N1型高致病性禽流感病毒可能是一个新的药物靶点。更多还原

【Abstract】 The NS1 protein, a non-structural protein of Avian Influenza Virus, is about 230 amino acids in length. Its molecular weight is about 26 KDa. The protein has two functional domains: an RNA binding domain in the amino terminal and an effector domain in the carboxyl terminal. As a multifunctional viral protein, the NS1 protein can not only influence the expression of other genes of the virus, but also counteract host defense system by interacting with many cellular proteins. Therefore, the NS1 protein is regarded as a crucial virulent factor for Avian Influenza Virus. To find other cellular proteins that interact with NS1, to explore new functions of NS1 and then to elucidate the possible mechanism of NS1 pathogenicity, a human thymus cDNA library was screened using yeast two-hybrid system and eight genes were found to interact with NS1. We chose U protein as our object protein and did yeast point to point back-hybrid to identify its interaction. We also found that U interacts with NS1 RNA binding domain. The interaction of these two proteins was further confirmed by GST pull down. From our study, we also found that the 80-84aa deletion of HPAIV NS1 may be very important for NS1 pathogenicity. This indicates a new drug target for lethal H5N1.更多还原