猪流行性腹泻病毒Nsp7的亚细胞定位和对Ⅰ型干扰素应答的影响

doi:10.11843/j.issn.0366-6964.2017.03.013

Abstract

Abstract:

To investigate subcellular localization and effect on type Ⅰ interferon (IFN) response of nonstructural protein 7 (Nsp7) of porcine epidemic diarrhea virus (PEDV), the Nsp7 gene sequence was predicted by bioinformatics, the Nsp7 gene of Chinese PEDV epidemic strain was cloned and inserted into the eukaryotic expression vector pCAGGS. The expression and subcellular localization of Nsp7 in transfected cells were determined by Western blot and indirect immunofluorescence assay respectively. The effect of Nsp7 on type Ⅰ IFN response was evaluated by dual luciferase reporter gene assay, ELISA and virus replication-inhibition bioassay, respectively. Gene cloning and sequence analysis results showed that PEDV Nsp7 gene was 249 bp in length, and it was highly conserved among different PEDV strains. Western blot and indirect immunofluorescence assay showed that Nsp7 was highly expressed and localized mainly in cytoplasm. Dual luciferase reporter gene assay indicated that Nsp7 strongly inhibited the IFN-β promoter activity. ELISA results showed that Nsp7 could significantly inhibit the expression of IFN-β in the protein level. VSV replication assay revealed that Nsp7 significantly inhibited type Ⅰ IFN antiviral activity induced by poly(I:C). Our results implied that Nsp7 was a highly conserved protein of PEDV and exhibited antagonistic function on type Ⅰ IFN response. The results have laid a foundation for exploring the innate immunity evasion mechanism of PEDV and guiding the clinical application of vaccines.

CLC Number:

S852.659.6

LI Hong-jie, WANG Xiao-xue, GAO Dong-sheng, HUANG Hui-min, CHEN Lu, CHANG Hong-tao, WANG Chuan-qing, LI Yong-tao, ZHAO Jun. Subcellular Localization and Effect on Type Ⅰ Interferon Response of Porcine Epidemic Diarrhea Virus Nsp7[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2017, 48(3): 501-507.

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Subcellular Localization and Effect on Type Ⅰ Interferon Response of Porcine Epidemic Diarrhea Virus Nsp7

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