Figure 2.

The carboxyl half of pUL37 is sufficient for replication of virus in tissue culture. Mutant viruses were analyzed by single-step growth curves. Vero cells (5 × 10⁵) were infected at MOI of 10 PFU/cell. The infected cells were harvested at 24 h post-infection. The lysates were sonicated and serial dilutions plated on BD45 cell monolayers to enumerate virus titer. Data are plotted as PFU/mL from the infected cell lysate with input virus inoculum indicated (A) and percent titer relative to the wild-type (WT) UL37–EGFP virus (B). Each value is an average of two biological replicates. (C) Accumulation of the pUL37 mutant polypeptides was examined by immunoblot methods. Vero cells were infected as above and total cell lysates were analyzed by SDS-PAGE (4–12% NuPAGE gradient gel), followed by transfer to nitrocellulose membranes. The blot was probed with rabbit anti-GFP antibodies to confirm stable expression of all truncated pUL37–EGFP fusion proteins. Protein standards are shown on the left of the blot.