靶向PRRSV核衣壳蛋白N基因的siRNA表达载体的构建及鉴定

Construction and Identification of siRNA Expression Vector Targeting Nucleocapsid Protein N Gene of PRRSV

[目的]研究靶向PRRSV核衣壳蛋白N基因的siRNA表达载体的构建及鉴定。[方法]设计并人工合成靶向PRRSV核蛋白N基因的3个siRNA靶序列,将其插入CMV启动子下游,克隆到真核表达载体pSilencer4.1-CMV中,对该重组表达载体进行酶切鉴定及DNA测序。[结果]成功构建了靶向核蛋白基因表达的siRNA干扰重组质粒载体pSilencer-N。[结论]该研究为进一步运用RNA干扰技术进行PRRSV防治研究奠定了基础。

[ Objective] The aim was to research the construction and identification of siRNA expression vector targeting nucleocapsid protein N gene of PRRSV. [ Method] According to sequenced nueleocapsid protein N gene sequence of PRRSV,three siRNA oligonueleotides targeting nucleoeapsid protein N gene were designed and synthesized,and were inserted into CMV promoter downstream. Oligonueleotides were inserted into pSileneer 4.1- CMV vector,these recombined plasmids were identified by restriction endonuelease and DNA sequencing. [ Result ] siRNA expression vector of nueleocapsid protein N gene has been successfully constructed. [ Conclusion ] This study lays a foundation for eontrolling PRRSV by RNA interference technique.