短发夹结构RNA对口蹄疫病毒2B基因瞬时表达的抑制  

Inhibition of food-and-mouth disease virus 2B gene transient expression by short hairpin structure RNA

作　　者：崔丽瑾[1] 王兴龙[2] 任林柱[1] 伍晓慧[3] 郎需龙[2] 张辉[1] 梅英武[1] 李晓燕[2] 卜朝阳[2] 

机构地区：[1]吉林大学畜牧兽医学院,吉林长春130062 [2]军事医学科学院军事兽医研究所,吉林长春130062 [3]河北农业大学海洋学院,河北秦皇岛066003

出　　处：《中国兽医学报》2008年第5期532-535,共4页Chinese Journal of Veterinary Science

基　　金：吉林省科技发展资助项目(20050549)

摘　　要：为探讨siRNA在哺乳动物细胞中对口蹄疫病毒(FMDV)2B基因的抑制作用,针对WFL株FMDV的2B基因,设计了4个siRNA,并将shRNA表达模板插入pSilencer1.0-U6载体,构建shRNA表达质粒,将其与含绿色荧光蛋白报告基因的pEGFP+2B融合表达质粒瞬时共转染BHK-21细胞,以荧光显微镜和流式细胞仪检测荧光表达量的变化,以实时定量RT-PCR法检测对2B基因mRNA的抑制作用。结果表明,本试验所构建的针对2B基因的shRNA表达载体可以在BHK-21细胞中抑制2B基因的瞬时表达。To investigate the interference of FMDV 2B gene expression in mammalian cells by small interfering RNAs （siRNAs）, four shRNAs expression plasmids targeting the 2B gene and the fusion expression plasmid of pEGFP 4-2B were constructed and transiently cotransfected into BHK-21 cells. The diversify of expression amount of EGFP was detected by fluorescence microscope and FCM,and the transcriptional level of 2B in BHK-21 cell was detected by real-time RT-PCR, The result showed that the 2B gene transient expression was inhibited by shRNA expression plasmids targeting the 2B gene which we recombined. It suggested that RNAi might represent a new approach for the treatment of FMDV.

关 键 词：FMDV RNA干扰 2B基因 

分 类 号：S852.65[农业科学—基础兽医学]