Abstract:

The eukaryotic expression vectors for the full-length and truncated forms of the mitochondrial antiviral-signaling protein (MAVS) was established and their expression in HEK293 cells was detected, which provides important experimental materials for subsequent investigations into the viral escape mechanism mediated by MAVS. Firstly, the full-length human MAVS sequence from the NCBI database was analyzed to design primers for generating truncated forms. Gene fragments corresponding to the full-length and various truncated forms of MAVS were amplified from cDNA templates using PCR and cloned into the pcDNA3.1-3xflag vector. Subsequently, the constructed vectors were transfected into HEK-293 cells. RT-PCR was employed to verify the size of the amplified products, which were compared with the target sizes, and Western blot analysis was used to detect the expression of MAVS proteins in the 293 cells. Eukaryotic expression vectors for full-length MAVS (MAVS-FL) and its truncates MAVS-△C, MAVS-△CP, MAVS-PBD, and MAVS-△Tm were successfully constructed. Western blot results confirmed that the proteins encoded by these vectors were effectively expressed in HEK-293 cells. In this study, we successfully constructed eukaryotic expression vectors for the full-length and truncated forms of MAVS and expressed these proteins in HEK-293 cells. This work provides important experimental materials for further study of the functions and antiviral mechanisms of MAVS proteins.

Key words: mitochondrial antiviral protein, eukaryotic expression vector, truncated forms construction, protein expression