Figure 2

Essentiality of A12L protein in VV replication. In order to determine the essentiality of A12L protein in virus replication, T-REx 293 cells were infected with vvtetO A12L in the presence/absence of Tet (Tet+/-). The lack of A12L was complemented by the transient expression of plasmid born A12L under the control of an early/late synthetic promoter (pA12L) or the native promoter (233 nucleotide upstream of A12L ORF, p233-A12L). In addition, the N-terminal AG/A site mutated A12L was constructed to rescue the absence of A12L (AG/A). pA12L: A12L ORF under the control of the early/late synthetic promoter; p233-A12L: plasmid born A12L under the native promoter; pRB21: vector plasmid alone; AG/A: plasmid born A12L with N-terminal AG/A site mutation into ID/I. Each virus titer (PFU/ml) was scaled in log phase.