FIG 4.

Mutant rVSV-MAK-GP T544I virus outcompetes wild-type virus in competition assays. (A) Schematic showing the workflow of the competition assay and time course collection of viral progeny in the P1 and P2 samples. Vero E6 cells were coinfected with increasing amounts of the wild-type virus and a constant amount of the T544I mutant virus (MOI of 0.001) at three different ratios, 1:1, 10:1, and 100:1. Supernatant virus samples were collected for sequencing at the indicated times. (B) Frequency of I544 and T544 residues in virus progeny from the coinfections described for panel A. cDNAs synthesized from viral genomes were sequenced by the Sanger method, and the percentages of genomes encoding T544 or I544 were calculated as described in Materials and Methods. Values of >99% indicates that only one amino acid was detected in the trace files.