H9N2亚型禽流感病毒PA-X蛋白功能研究  

Study on the Function of PA-X Protein of H9N2 Avian Infl uenza Virus

作　　者：李鲁兆 崔宏锐 林伟山 李雪松[1] 杨健美[1] 滕巧泱[1] 刘芹防[1] 李泽君[1] LI Luzhao;CUI Hongrui;LIN Weishan;LI Xuesong;YANG Jianmei;TENG Qiaoyang;LIU Qinfang;LI Zejun(Shanghai Veterinary Research Institute,CAAS,Shanghai 200241,China)

机构地区：[1]中国农业科学院上海兽医研究所,上海200241

出　　处：《中国动物传染病学报》2022年第4期13-19,共7页Chinese Journal of Animal Infectious Diseases

基　　金：“十三五”重点研发计划(2016YFD0500204);国家自然科学基金项目(31702237);上海市自然科学基金项目(17ZR1437400);上海科委创新行动计划农业项目(17391901700)。

摘　　要：本研究选取了一株H9N2亚型禽流感病毒A/Chicken/Shanghai/2093/2009毒株(简称A2093),分别构建了共表达PA-X和PA蛋白的真核表达质粒、单表达PA蛋白和PA-X蛋白的真核表达质粒,研究PA-X对宿主蛋白表达及流感病毒聚合酶活性的影响。结果显示:同时表达PA-X和PA蛋白可以强烈抑制宿主蛋白表达,单独表达A2093的PA-X同样具有较强抑制宿主蛋白表达的功能,而仅表达PA蛋白不影响宿主蛋白表达;在A2093的复制子系统中PA-X缺失后聚合酶活性明显提高;利用反向遗传操作技术拯救出缺失PA-X的A2093ΔPA-X毒株,在MDCK和LMH细胞上比较其与亲本A2093毒株复制能力的差异,发现在MDCK和LMH细胞上,从接种后12~36 h的不同时间点A2093ΔPA-X比A2093病毒低度明显降低。In this study,a strain of H9N2 avian influenza viruses A/Chicken/Shanghai/2093/2009(A2093)was used to construct the eukaryotic expression plasmids for co-expressing PA-X and PA or expressing PA and PA-X alone in order to investigate the infl uence of PA-X on host protein expression and polymerase activity.Either co-expression of PA-X and PA or expression of PA-X alone strongly inhibited the expression of host proteins while expression of PA alone did not.Higher polymerase activity was found in the mini-replicon system of A2093 when PA-X was deleted.The virus A2093ΔPA-X without PA-X was rescued using the reverse genetic technology and its replication capabilities on MDCK and LMH cells were compared with the parental virus A2093.The results showed that the virus titers of A2093ΔPA-X were lower than that of A2093 on both MDCK and LMH cells at different time points during 12 to 36 hours'observation.

关 键 词：H9N2亚型禽流感 PA-X 复制 功能 

分 类 号：S852.65[农业科学—基础兽医学]