Construction and Expression of Eukaryotic Expression Plasmid of FMDV 3D Gene and Its Effect on Type I IFN Signaling Pathway
Miao Qin
Wu Xiangju
Qi Jing
Cong Xiaoyan
Li Juntong
Wang Lin
Du Yijun
Abstract：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.
Keywords：Foot-and-mouth disease virus3D proteinEukaryotic expression plasmidType Ⅰ IFN
Publication Date：2024-02-28
Online Publishing Date：2026-05-22(First online date of this platform, not the publication date of the document)
Pages：7( 144-150 )