This study aimed to construct the eukaryotic expression plasmid of 3D gene of foot-and-mouth disease virus(FMDV)and explore its effects on type Ⅰ interferon(IFN)signaling pathway.The plasmid pXJ41-Myc-3D was constructed after 3D gene was synthesized according to the GenBank sequence and inserted into the eukaryotic expression vector pXJ41,which was transfected to HEK-293T and PK-15 cells after identi-fied by PCR,double digestion and nucleotide sequencing.The expression and cellular localization of 3D pro-tein were detected by Western blotting and indirect immunofluorescence assay(IFA).The effects of 3D over-expression in HEK-293T cells on type Ⅰ IFN signaling pathway induced by versicular stomatitis virus(VSV)were further assayed by dual luciferase reporter assays,Real-time PCR and TCID50.The results above showed that the eukaryotic expression plasmid pXJ41-Myc-3D was constructed successfully,the 3D protein could be expressed in HEK-293T cells with the size about 55 kDa and mainly localized in the nucleus,3D protein could inhibit the promoter activity and mRNA level of IFN-β induced by VSV and promote replications of the virus.This study laid a foundation for further exploring the mechanism of 3D protein inhibiting type Ⅰ IFN signaling pathway.
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