猪繁殖与呼吸综合征是由猪繁殖与呼吸综合征病毒（PRRSV）引起的以母猪繁殖障碍、仔猪死亡以及各年龄的猪呼吸道症状为特征的疾病，严重威胁着养猪业的健康发展。本课题组前期利用iTRAQ技术对PRRSV高致病性毒株感染的MARC-145细胞进行蛋白质组学分析，发现反义通路（antisense pathway）呈显著上调活化。在本项目中我们利用Real-time PCR、Western-blotting等技术分析PRRSV感染细胞后反义通路的活化情况，并利用链特异性转录组测序技术对病毒感染后细胞转录的正链mRNA和反义mRNA进行定量，确定反义通路活化对蛋白质合成的影响。通过对反义通路关键蛋白（PSF和p54nrb）进行过表达和抑制表达，分析该通路对病毒感染的影响，然后利用免疫共沉淀和激光共聚焦等技术分析病毒入核蛋白与反义通路关键蛋白的相互作用，阐明反义通路与PRRSV相互作用的分子机制。

Porcine reproductive and respiratory syndrome caused by porcine reproductive and respiratory syndrome virus (PRRSV) can lead to reproductive failure in sows, death in piglets and respiratory symptoms of swine in all ages, which is a serious threat to the healthy development of pig industry. We carried out proteomic analysis of MARC-145 cells infected with highly pathogenic PRRSV strain using iTRAQ technology, and found that antisense pathway was activated and significantly up-regulated. In this project we analyze the activation of antisense pathway in PRRSV-infected cells using real-time PCR, western-blotting and so on, and quantify the positive strand mRNA and antisense mRNA during cellular transcription after viral infection to determine the effect of the activation of antisense pathway on protein synthesis using strand specific transcriptome sequencing technique. Based on the over-expression and inhibited expression of antisense key proteins (PSF and p54nrb), we confirm the impact of antisense pathway on viral infection, then explore the interaction between viral proteins distributed in cell nucleus and key proteins (PSF and p54nrb) of antisense pathway using co-immunoprecipitation and confocal laser techniques to elucidate the molecular mechanism of interaction between antisense pathway and PRRSV.