Nsp9基因在Marc-145细胞上对猪繁殖与呼吸综合征病毒复制的影响

Effect of Nsp9 Gene on Replication of PRRSV in Marc-145 Cells

试验旨在验证Nsp9基因是否影响猪繁殖与呼吸综合征病毒(porcine reproductive and respiratory syndrome virus,PRRSV)的复制。构建含有Nsp9全长基因的质粒pIRES2-EGFP-Nsp9,并转染到Marc-145细胞中,接毒之后分别通过实时荧光定量PCR和Western blotting方法测定PRRSV N蛋白在mRNA和蛋白水平的表达。结果显示,转染Nsp9基因的Marc-145细胞上PRRSV的N蛋白在mRNA水平上显著高于未转染Nsp9基因的对照组(P<0.05),是对照组的1.5倍,同时转染Nsp9基因后的Marc-145上PRRSV N蛋白水平也明显高于对照组。随着剂量的增加,N蛋白的表达量在mRNA和蛋白水平都有所增加。由以上结果初步可知,Nsp9基因可以在Marc-145细胞上促进PRRSV的复制,Nsp9基因与其复制密切相关。

In order to evaluate whether Nsp9 could enhance the replication of porcine reproductive and respiratory syndrome virus （PRRSV）,pIRES2-EGFP-Nsp9 plasmid containing whole Nsp9 genome were transfected into Marc-145 cells,Real-time PCR and Western blotting were used to evaluate the expression of N protein after PRRSV was inoculated.The results showed that the level of N protein in the Nsp9 transfected cells was significantly higher than control group （P 〈0.05）,which was 1.5 times higher.Meanwhile,the expression at protein level had the same trend as the mRNA level.When the dose of plasmid was increasing,the expression of N protein both at the mRNA and protein levels were increased.In conclusion,Nsp 9 gene could enhance the replica-tion of PRRSV in Marc-145 cells.