Figure 2: Generation of recombinant vesicular stomatitis virus (rVSV) vaccines. The glycoprotein (G) gene in rVSVwt was deleted and replaced by the Ebola virus glycoprotein (EBOV-GP) resulting in the vector rVSV/EBOV-GP. Co-cultures of 293T and Vero cells were transfected with the following plasmids: full-length genome plasmid rVSVΔG/EBOV-GP expressing EBOV-GP under control of the bacteriophage T7 polymerase promoter; plasmids expressing the proteins needed for VSV replication, nucleoprotein (N), phosphoprotein (P) and polymerase (L) under control of the human cytomegalovirus CMV promoter; and the plasmid expressing the bacteriophage T7 polymerase under control of the CMV promoter. Following incubation of 24-72 hrs rVSV/EBOV-GP particles are harvested and purified from the cell supernatant.