| 摘要: |
| 【目的】研究猪流行性腹泻病毒（PEDV）M蛋白对Ⅰ型干扰素信号通路的影响。【方法】克隆PEDV M基因，构建其真核表达载体pcDNA3.1-Myc-PEDVM，并转染HEK-293T细胞。采用Western bloting试验检测M蛋白的表达。通过双荧光素酶报告试验检测M蛋白对SeV诱导的干扰素通路活化的影响；通过Real-time PCR分析M蛋白对SeV诱导的IFN-β及干扰素刺激基因ISG54、ISG56、RIG-I mRNA表达的影响。通过过表达试验验证上调表达M蛋白对PEDV复制的影响。【结果】克隆了676 bp的PEDV M基因，成功构建其真核表达载体pcDNA3.1-Myc-PEDV-M，且其可在HEK-293T细胞中表达。PEDV M蛋白可抑制Ⅰ型干扰素信号通路的激活，且M蛋白对SeV诱导的Ⅰ型干扰素通路活化的抑制具有剂量依赖效应；M蛋白能够抑制IFN-β与干扰素诱导基因ISG52、ISG56、RIG-I mRNA的表达，并具有剂量依赖效应。过表达试验表明，M蛋白在抑制Ⅰ型干扰素信号通路激活的同时促进PEDV复制。【结论】PEDV M蛋白能够抑制Ⅰ型干扰素信号通路的激活，并促进PEDV复制。 |
| 关键词: 猪流行性腹泻病毒 PEDV M蛋白 干扰素信号通路 |
| DOI： |
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| 基金项目:国家科技计划(科技支撑-子课题)(2015BAD12B00-2)；四川省科技支撑计划项目(2017NZ0038，2016NYZ0052） |
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| Inhibitory effect of PEDV M protein on type Ⅰ interferon signaling pathway |
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CHEN Pan,LI Shasha,ZHU Zixiang,et al
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| Abstract: |
| 【Objective】This study investigated the effect of porcine epidemic diarrhea virus (PEDV) M protein on type Ⅰ interferon (IFN) pathway.【Method】PEDV M gene was cloned to construct its eukaryotic expression vector pcDNA3.1-Myc-PEDV-M,and the expression of M protein was verified by Western bloting analysis.A dual luciferase reporter assay was performed to evaluate the effect of M protein on SeV induced type Ⅰ IFN pathway activation.The inhibitive effect of M protein on the mRNA of IFN-β,ISG54,ISG56,and RIG-I was measured by qPCR analysis.The overexpression assay was further performed by transfection of the plasmids expressing M protein into porcine cells to investigate the effect of M protein on PEDV replication.【Result】The PEDV M gene of 676 bp was cloned,its PEDV M eukaryotic expressing plasmid was successfully constructed and the expression of M protein in HEK-293T was verified.The dual luciferase reporter assay demonstrated that PEDV M protein inhibited type Ⅰ IFN pathway signal transduction in a dose dependent manner,and M protein significantly impaired SeV induced IFN-β,ISG52,ISG56,and RIG-I expression with dose-dependent effect.Overexpression of M protein promoted PEDV replication and suppressed type Ⅰ IFN pathway signal transduction.【Conclusion】PEDV M protein inhibited type Ⅰ IFN pathway signal transduction and promoted PEDV viral replication. |
| Key words: porcine epidemic diarrhea virus PEDV M protein interferon pathway |
