siRNA对乙型脑炎病毒复制的抑制效果  

Inhibition of Japanese Encephalitis Replication in Cells by Small Interfering RNAs

作　　者：刘学芹[1,2] 曹胜波[1,2] 李么明[1,2] 孙敏轩[1,2] 叶静[1,2] 周锐[1,2] 杨影[1,2] 徐高原[1,2] 陈焕春[1,2] 

机构地区：[1]华中农业大学农业微生物学国家重点实验室,武汉430070 [2]华中农业大学水产学院,武汉430070

出　　处：《华中农业大学学报》2009年第2期188-192,共5页Journal of Huazhong Agricultural University

基　　金：国家自然科学基金项目(30600446);湖北省重大科技攻关项目(2006AA205A02)资助

摘　　要：以JEV的NS1基因mRNA为靶序列,设计合成了4个siRNA序列,构建了各自的表达质粒pS-NS1A、pS-NS1B、pS-NS1C和pS-NS1D。通过间接免疫荧光、Western blot检测、RT-PCR和病毒空斑检测等方法对这些siRNAs抑制JEV复制的效果进行了评价。结果表明4个siRNA表达质粒均对JEV在细胞中的复制具有不同程度的抑制作用,其中pS-NS1C、pS-NS1D有显著的抑制作用。说明针对NS1基因的siRNAs可以有效抑制JEV的复制,并有望成为应对JEV感染的治疗方法。To determine whether Japanese encephalitis virus（JEV） replication in cell lines can be inhibited by small interfering RNAs（siRNAs） which targets NS1 gene, four siRNAs expression plasmids were generated to target different sites of the NS1 gene. JEV replication and viral protein expression were evaluated by indirect immunofluorescence assay,Western blot,RT-PCR and viral titration. The results showed that JEV replication was inhibited by these siRNAs in BHK-21 cell line with different efficiency. Two of siRNAs plasmids, pS-NS1C and pS-NS1D,were demonstrated as the most efficient siRNAs. Our results suggested that NS1 can be as a target gene for siRNAs to inhibit JEV replication and siRNAs can be used as potential therapy strategy for JEV infection in the future.

关 键 词：乙型脑炎病毒 NS1 siRNA复制 抑制 

分 类 号：S852.65[农业科学—基础兽医学]