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
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
作者信息
- 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
- 折叠
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cDNA,PCR扩增出猪的ESD基因,构建到pcDNATM3.1/Myc-HisA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.
Abstract
[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 cDNA and PCR amplification and constructed to pcDNATM3.1/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Key words
Foot and mouth disease virus/Esterase D/Antiviral effect引用本文复制引用
基金项目
国家自然科学基金(31502042)
国家自然科学基金(31302118)
科技支撑计划项目(2015BAD12B04)
甘肃省杰出青年基金(145RJDA328)
农业部948项目(2015-Z6)
出版年
2016
国家科技期刊平台
NSTL
维普
万方数据