Abstract:

This study focused on the effect of Rotavirus (RV) non-structural protein 5 (NSP5) on RV replication in vitro. After the construction of eukaryotic expression vector for expressing recombinant protein NSP5 specifically based on pEGFP-N2, the recombinant plasmid was transfected into MA104 cells. The expression of NSP5 at different time was detected by Immunofluorescence and Western Blot respectively. Subsequently, the RV replication and the migration of NSP5 based on the distribution of GFP were confirmed by fluorescence observation when MA104 cells were transfected with recombinant plasmid and empty plasmid infected with the same dose of RV. The recombinant plasmid expressed the strongest at 48 h post transfection in MA104 cells. Moreover, the location of recombinant protein NSP5 changed from dispersion to aggregation because of RV infection, and RV replication in the cells transfected with pEGFP-N2-NSP5 was significantly higher than in the control cells. In this study, the NSP5 could promote RV replication, which provides theoretical basis for further study on RV infection, replication and pathogenesis.

Key words: rotavirus, NSP5, eukaryotic expression vector, virus replication