Inhibition Of Porcine Reproductive And Respiratory Syndrome Virus Non-structural Protein1αand11Expression By Small Interfering RNA

Porcine reproductive and respiratory syndrome, which was caused bythe virus of porcine reproductive and respiratory syndrome and also known asblue-ear disease, is a continuous or intermittent emergence severe illness. ORF1a andORF1b of PRRSV encode respectively which are both non-structural aggregationprotein. As pp1ab and pp1a were synthesized by genomic mRNA, pp1ab and pp1awere cleaved into14non-structural proteins with different functions by serineproteases,3c-cysteine proteinase and PCP which were all coded by ORF1a. Such as,nsp1α can inhibit IB phosphorylation, thereby inhibit activation of NF-κB gene,resulted in inactivation of NF-κb,then prevent the generation of IFN-β. Nsp11canregulate RIG-1-mediated signaling pathways by the triggering of dsRNA to proventthe production of IFN-β.RNA interference (RNA interference, RNAi) was a specific silencing mechanismwhich was initiated by a double-stranded RNA,it can inhibit or turn off the expressionof genes related sequences.In recent years,with the development of molecular biology,RNAi technology has been widely applied in various fields prevention, such as cancer,gene function and studies of animal diseases. siRNA, endonuclease and otherproteins,were formed a RISC (RNA-induced silencing complex), among them, partof nucleic acid can lead protein to break down mRNA.In this study，Synthetic siRNA of nonstructural proteins nsp1α and nsp11ofPRRSV，siRNA which was with recombinant plasmid pcDNA3.1-nsp1α-GFP，pcDNA3.1-nsp1α-GFP co-transfected293T cells and MARC-145cells respectively.By fluorescence microscopy, Western Blot, indirect immunofluorescence test and measurement of virus TCID, the effects of siRNA to non-structural proteins of nsp1αand nsp11were revealed.