Fig. 2.

(A) Schematic of the inserts for Late Venus (LV), Intermediate Venus (IV), Early Venus (EV), and Promoter-less Venus (PLV) single reporter viruses. Inserts were placed between the Vaccinia J4R and J5L genes, as shown in Fig. 1. The canonical late F17R, intermediate G8R, and early C11R viral promoters were used. EV contains an early terminator sequence (term) immediately after the Venus stop codon. PLV contains the Venus open reading frame with no preceding promoter element. (B) Kinetics of Venus fluorescence following high MOI infection of A549 cells with either LV, IV, EV, or PLV. Raw fluorescence is plotted in log-scale. (C–E) Effect of the DNA replication inhibitor AraC on reporter expression following EV infection (C), IV infection (D), or LV infection (E), in the same experiment as (B). Open circles are untreated infections and filled circles are infections in the presence of 1 μg/ml AraC. S/B ratios are plotted.