外泌体源性oar-miR-10b在绵羊痘病毒感染绵羊睾丸细胞中的动态表达及靶基因的鉴定  

Dynamic Expression of Exosome-derived Oar-miR-10b after the Ovine Testicular Cells Infected with Sheep Pox Virus and Identification of Its Target Genes

作　　者：孔贺磊 吴金恩[1] 郑亚东[2] 何贵天 李雅婷 丁军涛[1] KONG He-lei;WU Jin-en;ZHENG Ya-dong;HE Gui-tian;LI Ya-ting;DING Jun-tao(College of Life Science and Technology,Xinjiang University,Urumqi,Xinjiang,830046,China;State Key Laboratory of Veterinary Etiological Biology/Lanzhou Veterinary Research Institute,Chinese Academy of Agricultural Sciences,Lanzhou,Gansu,730046,China)

机构地区：[1]新疆大学生命科学与技术学院,新疆乌鲁木齐830046 [2]中国农业科学院兰州兽医研究所/家畜疫病病原生物学国家重点实验室,甘肃兰州730046

出　　处：《动物医学进展》2018年第12期42-48,共7页Progress In Veterinary Medicine

基　　金：新疆维吾尔自治区自然科学基金(2016D01C041)

摘　　要：以绵羊痘病毒(SPPV)感染绵羊睾丸细胞4h后外泌体中miRNAs的高通量测序结果为基础,应用RT-qPCR分析SPPV感染绵羊睾丸细胞4h、24h、48h和72h后外泌体中oar-miR-10b的动态表达;用双荧光素酶报告载体鉴定oar-miR-10b的靶基因。结果显示,在SPPV感染的不同时期,oar-miR-10b呈先增加后减少的趋势,特别是感染后24h的差异表达量为2.15倍;PmirGLO-BCL2L2-WT+oar-miR-10b mimics共转染组与PmirGLO-BCL2L2-WT+miRNA Negative Control对照组相比差异极显著(P <0.01),证实BCL2L2为oar-miR-10b的靶基因。结果表明,oar-miR-10b在SPPV感染的不同时期呈差异性表达,其靶基因为BCL2L2,为深入研究oar-miR-10b在SPPV感染中的作用机制奠定基础。In order to study the dynamic expression of oar-miR-10b in ovine testicular cells infected with SPPV,and identify its target genes,based on the results of high-throughput sequencing of miRNAs in exosomes of SPPV-infected ovine testicular cells after4h,dynamic expression of oar-miR-10b in exosomes was detected by RT-qPCR at4h,24h,48h and72h after SPPV infection in ovine testicular cells.The interaction between oar-mir-10b and target genes was identified by dual luciferase report system.The results showed that Oar-miR-10b in the exosomes was differentially expressed in the different stages of SPPV-infected ovine testicular cells,which showed a tendency to increase first and then decrease,the differential expression was to2.15times after24h infection,especially.PmirGLO-BCL2L2-WT+oar-miR-10b mimic co-transfection group was significantly different from PmirGLO-BCL2L2-WT+miRNA negative control co-transfection control group,this result indicated that the target gene of oar-miR-10b is BCL2L2.This study preliminary confirmed that oar-miR-10b in the different stages of the infection of SPPV is differentially expressed and its target gene is BCL2L2,which lays the foundation for further study on the mechanism of oar-miR-10b in the process of SPPV infection.

关 键 词：oar-miR-10b BCL2L2 实时荧光定量PCR 双荧光素酶报告系统 

分 类 号：S852.659.1[农业科学—基础兽医学]