针对nsp9基因的siRNA在Marc-145细胞上对猪繁殖与呼吸综合征病毒复制的影响

Influence of siRNA targeting nsp9 gene on replication of PRRSV in Marc-145 cells

为了验证猪繁殖与呼吸综合征病毒(PRRSV)nsp9基因对PRRSV的复制影响,化学合成针对nsp9基因的siRNA,转染到Marc-145细胞中,接种PRRSV之后分别通过荧光定量和Western blot方法来测定病毒的滴度变化和表达情况。结果发现,转染siRNA的Marc-145细胞中病毒的滴度低于未转染特定siRNA的对照组,同时N蛋白的mRNA和蛋白水平均低于未转染特定siRNA的对照组。初步得出结论nsp9基因为病毒复制关键基因,针对nsp9基因的siRNA会在Marc-145细胞中抑制PRRSV的复制。

In order to verify whether nsp9 gene of PRRSV have an influence on the replication of PRRSV,chemical synthesis siRNA special for nsp9 gene was transfected into Marc-145 cells, qPCR and Western blot were conducted to evaluate viral titer and expression after inoculated PRRSV, The results showed that viral titer in the Marc-145 ceils transfected with siRNA were lower than that of not transfected cells. Meanwhile,the mRNA and protein level were the same as the titer level. So nsp9 is a crucial gene for viral replication. SiRNA for nsp9 gene could inhibit the replication of PRRSV in Marc-145 cells.