Figure 2.

Viral protein requirements for plasmid replication. (A) Conditional lethal ts mutants. BS-C-40 cells were transfected with p716 and 24 h later were mock infected or infected with 3 PFU per cell of wild type VAC (WR) or ts mutant Cts24, Cts25, ts185, or Cts16 at permissive (31°C) and non-permissive (39.5°C) temperatures for 24 h. DNA was then isolated, digested with DpnI and quantified by real-time PCR. (B) D4R deletion mutant. RKD4R and RK-13 cells were transfected with p716 and 24 h later were infected with 3 PFU of vD4-ZG. At 24 after infection, DNA was isolated, digested with DpnI, and the amount of DNA quantified by real-time PCR.