The generation of plasmid RGV14 for use in studying the effect of F1-R fusion protein mutations on enhanced cleavability of Sendai virus

Wild type (wt) Sendai virus (SeV), a negative sense RNA Paramyxovirus, causes respiratory infections in rodents. It is a model for respiratory tract infections in humans caused by other Paramyxovirdae viruses. F1-R, a Z-strain variant, causes a systemic (pantropic) infection, which has been correlated with two phenotypes not seen in wt SeV. These are enhanced cleavability of the Fusion protein (F) and budding from both the apical and basolateral domains of polarized cells. Previous studies have shown enhanced cleavability of the Fusion protein (F) requires the mutation F115 and one or more additional mutations in F1-R F. Mutation F104, which results in a loss of an N-glycosylation site, has been shown to confer enhanced fusion activity. To determine if F104 mutation contributes to the enhanced cleavability of F, plasmid RGV14 (Reverse Genetics Virus 14) containing mutations encoding F115 and F104 was generated. However, viral recovery was unsuccessful. Future studies will use an optimized plasmid from which virus has successfully been recovered