基于RNA-Seq筛选新型鸭细小病毒感染鸭胚成纤维细胞的差异表达基因  

Application of RNA-Seq Technology for Screening Differentially Expressed Genes in Duck Embryo Fibroblast Cells Affected by Duck Parvovirus

作　　者：崔元[1] 李晓轩 孙继国[1,2] 袁万哲 Cui Yuan;Li Xiaoxuan;Sun Jiguo;Yuan Wanzhe(College of Animal Medicine,Agricultural University of Hebei,Hebei Baoding 071001;Hebei Engineering and Technology Research Center of Veterinary Biotechnology,Hebei Baoding 071001)

机构地区：[1]河北农业大学动物医学院,河北保定071001 [2]河北省兽医生物技术工程技术研究中心,河北保定071001

出　　处：《现代畜牧兽医》2018年第5期1-6,共6页Modern Journal of Animal Husbandry and Veterinary Medicine

基　　金：河北省研究生创新资助项目

摘　　要：为筛选鸭细小病毒(DPV)感染鸭胚成纤维(DEF)细胞后的差异表达基因,本研究利用DPV接种DEF细胞作为试验组,对照组加入相同体积的2%DMEM,培养至72 h后提取2组样品的总RNA,利用RNA-Seq技术筛选出DEF细胞感染DPV后的差异表达基因,并对其进行生物信息学GO功能分类和KEGG分析。结果显示,差异表达水平变化倍数(fold change,FC)在2倍以上的基因共有4 073个,其中上调表达基因1 904个,下调表达2 169个。GO功能分析显示,这些基因主要涉及到炎症反应、免疫反应、蛋白结合、转运活动等功能。KEGG信号通路分析表明,这些差异表达基因参与细胞因子受体相互作用、新陈代谢、TNF、NF-κB、Jak-STAT、Toll样受体等信号通路。应用荧光定量PCR(Real-time FQ-PCR)验证部分差异表达基因的检测结果显示,差异基因的相对表达量与RNA-Seq技术测序结果基本一致,表明了RNA-Seq检测结果的可靠性。本研究为进一步研究DPV的致病作用及宿主抗病机制奠定了基础。In order to screen the differentially expressed genes of duck embryo fibroblast cells which were infected by duck parvovirus(DPV),duck embryo fibroblast cells were inoculated with DPV culture supernatants while the control group of DEF cells were inoculated with 2%DMEM,and cultured for 72 h.Then the DPV infected DEF cells group and control group were extracted total RNA for screening the differentially expressed genes by RNA-Seq technology.The results showed that DPV infected group compared with control group,a total of 4 073 genes were differentially expressed in the levels more than two times,including 1 904 up-regulated genes and 2 169 down-regulated genes.Moreover,the GO analysis results indicated that the differentially expressed genes were related to inflammatory response,immune response,protein binding,transporter activity,etc.KEGG Pathway analysis results revealed that the differentially expressed genes were related to Cytokine-cytokine receptor interaction,metabolism,TNF,NF-κB,Jak-STAT,Toll-like receptor signaling Pathways,etc.In addition,expressions of ten differentially expressed genes were verified by real-time PCR,which confirmed that mRNA expression levels were in line with the results identified by RNA-Seq technology.This study laid a foundation for the further study of the pathogenic of DPV and the host disease resistance mechanism.

关 键 词：鸭细小病毒 鸭胚成纤维细胞 转录组测序 差异表达基因 

分 类 号：S852.61[农业科学—基础兽医学]