FIG. 1.

Generation of VSV-FAST. (A) The p14 FAST gene from a fusogenic reptilian reovirus was subcloned into the pVSV-XN genome plasmid between the glycoprotein (G) and polymerase (L) as EGFP had been inserted previously. The G-deleted version was then made by replacing G with RFP. (B) Western blot analysis of infected cell lysates and sucrose banded virus reveals that FAST is expressed in VSV/FAST-infected cells and is present in the viral particle. Vero cells were infected with virus overnight, and cellular lysates were run (lanes 1 to 4) or virus was banded on a sucrose gradient and 10⁸ PFU were run per lane (lanes 5 to 7). Blots were probed for VSV proteins (upper), FAST protein (middle), or GFP (bottom). Lanes: 1, VSV/GFP; 2, mock; 3, VSV/FAST; 4, ΔM51-VSV/FAST; 5, VSV/GFP; 6, VSV/FAST; 7, ΔM51-VSV/FAST. (C) Confluent Vero cells were infected with the indicated viruses, and photomicrographs taken 24 h later show the large multinucleated syncytia induced by the FAST viruses (arrows). Magnification, ×8.