激活干扰素β信号通路鸭坦布苏病毒结构蛋白的鉴定  

Identification of duck Tembusu virus structural proteins activating interferonβsignaling pathway

作　　者：贺言 李国红 商雨 温国元 张蓉蓉 曾驰[1] HE Yan;LI Guo-hong;SHANG Yu;WEN Guo-yuan;ZHANG Rong-rong;ZENG Chi(School of Life Science and Technology,Wuhan Polytechnic University,Wuhan 430023,China;Key Laboratory of Prevention and Control Agents for Animal Bacteriosis(Ministry of Agriculture),Institute of Animal Husbandry and Veterinary,Hubei Academy of Agricultural Sciences,Wuhan 430064,China;Wuhan Keqian Biology Co.Ltd.,Wuhan 430070,China)

机构地区：[1]武汉轻工大学生命科学与技术学院,湖北武汉430023 [2]湖北省农业科学院畜牧兽医研究所农业农村部畜禽细菌病防治制剂创制重点实验室,湖北武汉430064 [3]武汉科前生物股份有限公司,湖北武汉430070

出　　处：《中国兽医科学》2022年第3期333-338,共6页Chinese Veterinary Science

基　　金：国家重点研发计划项目(2016YFD0500107);湖北省农业科技创新中心项目(2019-620-000-001-017);湖北省技术创新重大项目(2020ABA016)。

摘　　要：为了初步探究鸭坦布苏病毒(DTMUV)介导的鸭体天然免疫反应分子机制,本研究首先筛选出能显著激活IFN-β的DTMUV毒株,并将DTMUV的3个结构蛋白(C、PrM、E)克隆至真核表达载体pCAGGSHA,再分别与鸭源IFN-β启动子的双荧光素酶报告质粒duIFN-β-Luc、内参质粒pRL-TK共转染DEFs,使其在细胞内过表达,通过双荧光素酶报告试验检测DTMUV这3个结构蛋白激活鸭源IFN-β启动子的能力。结果显示,只有囊膜蛋白能显著激活IFN-β启动子,Western-blot试验证实这3个结构蛋白转染至DEFs均能正常表达。本研究结果表明,在DEFs中DTMUV的结构蛋白仅囊膜蛋白在激活机体天然免疫反应方面发挥了一定的作用。In order to preliminarily investigate the function of DTMUV on innate immune signaling pathway,at first it is proved that DTMUV infection does indeed trigger the activation of IFN-βpromoter in the DEFs,and the three structural proteins(C,Pr M and E)of DTMUV were cloned into eukaryotic expression vector p CAGGS-HA.Then the recombinant expression plasmid was contransfected into DEF cells with the duck IFN-βpromoter-luciferase reporter plasmid and the internal reference plasmid p RL-TK.IFN-βpromoter activity was determined by dual luciferase reporting system detection kit after transfection.As a result,that only overexpression of envelope significantly activated the IFN-βpromoter activity.Western-blot showed that three structural proteins of DTMUV was normally expressed in transfected DEF cells.Our study suggested that only the envelope of DTMUV’s structural proteins play a role in the activation of IFN-βsignaling pathway in DEF cells.

关 键 词：鸭坦布苏病毒 结构蛋白 双荧光素酶报告试验 IFN-Β 

分 类 号：S852.659.6[农业科学—基础兽医学]