靶向口蹄疫病毒IRES区RNA干扰位点的选择及shRNA表达载体的构建与鉴定

Screening of RNA Interfering Sites Targeting FMDV IRES Region and Establishment of shRNA Eukaryotic Expression Vectors

以口蹄疫病毒WFL株为试验株,对口蹄疫病毒的IRES序列进行了保守性分析和RNA二级结构预测,推测高度保守的FMDV复制原点区域位于容易被siRNA结合的IRESRNA二级结构的环区,因此,在该区选择确定了2个siRNA干扰靶位,设计并合成能表达其小发夹结构shRNA的表达模板,克隆到含有U6启动子的真核表达载体中,建立了靶向口蹄疫病毒IRES区的siRNA真核表达系统,为进一步研究针对该区的RNA干扰对口蹄疫病毒的抑制作用打下了基础。

Conservatism analysis and RNA secondary structure prediction of FMDV IRES identified that the highly conservative region of FMDV replication origin was located in the loop region of IRES RNA secondary structure which could be conjugated by siRNA easily,thus two siRNA sites were selected in this region and their shRNA expressing templates were designed and synthetized.Finally the templates were cloned into eukaryotic expressing plasmids which containing U6 promoters.This siRNA expression system may have a potential in the inhibition of FMDV replication.