Xu Z, Li X, Liu R, Si Y, Sun M, Jin M, Chen H, Qian P
College of Veterinary Medicine, Huazhong Agricultural University, Wuhan, Hubei, China.
Acta Virol. 2007;51(3):195-201.
The RNA-dependent RNA polymerase (RdRp) in viruses of the family Flaviviridae plays an important role in the viral replication process and in the forming of a replicase complex. We used small interfering RNAs (siRNAs) corresponding to the highly conservative Motif V of RdRp gene of different viruses to examine their role in modulating the expression of RdRp. Evaluation of the expression of RdRps was performed by the fluorescence, flow cytometry, Western blotting, and real-time PCR. We found that Classical swine fever virus (CSFV) siRNA could completely block the transcription and expression of RdRp. Additionally, Hepatitis C virus (HCV) siRNA could cause effective inhibition of RdRp, whereas Japanese encephalitis virus siRNA did not show significant repression of corresponding RdRp. These results demonstrated that siRNAs inhibited the expression of tested RdRps at the transcription level or at the posttranscriptional processing to a different extent.
黄病毒科病毒中的RNA依赖性RNA聚合酶(RdRp)在病毒复制过程以及复制酶复合体的形成中发挥着重要作用。我们使用了与不同病毒RdRp基因高度保守的基序V相对应的小干扰RNA(siRNA),来研究它们在调节RdRp表达中的作用。通过荧光、流式细胞术、蛋白质免疫印迹和实时PCR对RdRps的表达进行评估。我们发现,猪瘟病毒(CSFV)siRNA可完全阻断RdRp的转录和表达。此外,丙型肝炎病毒(HCV)siRNA可有效抑制RdRp,而日本脑炎病毒siRNA对相应的RdRp没有明显的抑制作用。这些结果表明,siRNA在转录水平或转录后加工阶段对所检测的RdRps表达有不同程度的抑制作用。