FIG. 8.

De novo expression of pUL21a is necessary and sufficient for efficient viral DNA synthesis and late accumulation of viral proteins. (A and B) Viral DNA synthesis (A) and protein accumulation (B) in GFP- or GFP/UL21a-expressing cells infected with recombinant virus that is generated in noncomplementing cells. MRC5-GFP cells or MRC5-GFP/UL21a cells were infected with ADwt and ADsubUL21a viruses at an input genome number equivalent to 2 TCID₅₀ units of wild-type virus/cell. Infected cells were collected at the indicated times and analyzed for viral DNA by qPCR (A) and for viral proteins by immunoblotting (B). (C) Viral protein accumulation in normal MRC-5 cells infected with recombinant virus that is generated in GFP- or GFP/UL21a-expressing cells. Normal MRC-5 cells were infected with ADwt or ADsubUL21a virus that was produced from either GFP (MRC5-GFP)- or GFP/UL21a (MRC5-GFP/UL21a)-expressing cells at an input genome number equivalent to 2 TCID₅₀ units of wild-type virus/cell. Infected cells were collected at the indicated times and analyzed for viral proteins by immunoblotting. Shown are representative results from at least two independent experiments. Error bars indicate standard deviations.