猪ESD真核表达质粒的构建及其抑制口蹄疫病毒复制的研究  

Construction of Porcine ESD Eukaryotic Expression Plasmid and Its Function to Suppress the Replication of Foot and Mouth Disease Virus

作　　者：李伟伟[1] 朱紫祥[1] 曹伟军[1] 杨帆[1] 李丹[1] 张克山[1] 刘湘涛[1] 郑海学[1] 

机构地区：[1]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,国家口蹄疫参考实验室,甘肃兰州730046

出　　处：《安徽农业科学》2016年第12期117-120,共4页Journal of Anhui Agricultural Sciences

基　　金：国家自然科学基金项目(31502042,31302118);科技支撑计划项目(2015BAD12B04);甘肃省杰出青年基金项目(145RJDA328);农业部948项目(2015-Z6)

摘　　要：[目的]探究猪ESD蛋白的抗病毒功能,通过构建猪ESD真核表达质粒,研究其是否能够抑制口蹄疫病毒在PK-15细胞中的复制。[方法]从PK-15细胞中提取总RNA,反转录为c DNA,PCR扩增出猪的ESD基因,构建到pc DNATM3.1/Myc-His A真核表达载体上。通过Western blotting和实时定量PCR检测ESD蛋白的表达情况及其抗病毒功能。[结果]获得了猪ESD真核表达载体,转染后能够正常表达。FMDV感染PK-15细胞后,ESD转录水平显著上调;过表达ESD蛋白显著抑制FMDV的复制;下调表达ESD蛋白显著促进FMDV的复制。[结论]猪ESD蛋白能够抑制FMDV在PK-15细胞中的复制。[Objective] To explore the antiviral effect of ESD protein,we constructed porcine ESD eukaryotic expression plasmid to investigate whether ESD protein to suppress the replication of foot and mouth disease virus in PK-15 cells.[Method] We extracted porcine cellular RNA from PK-15 cells,then porcine ESD gene was obtained by reverse transcription of c DNA and PCR amplification and constructed to pc DNATM3.1 / Myc-His A eukaryotic expression vector,of which expression and antiviral function were detected by western blotting and real-time quantitative PCR.[Result] The porcine ESD eukaryotic expression plasmid was constructed and ESD was successfully expressed.The mRNA level of ESD in PK-15 cells was up-regulated after FMDV infection.Overexpression of ESD significantly suppresses the replication of FMDV.Down-expression of ESD significantly promotes the replication of FMDV.[Conclusion] Porcine ESD protein significantly suppresses the replication of FMDV in PK-15 cells.

关 键 词：口蹄疫病毒 酯酶D 抗病毒功能 

分 类 号：S852.651[农业科学—基础兽医学]