Figure 1.

Constructs for VZV reporter viruses. The diagram depicts the presence of the recombinant sequences used to alter pOka [14] with BAC recombineering, as detailed in the text. Viruses shown in (A) include parent Oka and VZV-BAC-Luc, derived from the BAC-Zhu pOka system [14,19]. VZV-BAC-Luc was generated from pOka (passage 9), as previously described [19], and included a small population of TK-virus, which arose through unknown mechanisms. (B)—upper section: viruses derived from an uncorrected BAC system, as detailed by the Osterreider group, which include VZV-ORF9-T2A-Luc; VZV-ORF14-T2A-Luc; and VZV-ORF17-T2A-Luc. (B)—lower section: diagram of the viral protein and luciferase expression mediated by the T2A ribosome skipping motif, which is used to generate two proteins from one mRNA. (C) VZV-ORF57-T2A-Luc [27], VZV-ORF57-∆TK, VZV-ORF57-∆TS, and VZV-ORF57-∆TK∆TS were generated in a BAC with corrected spurious mutations identified in ORF40 and ORF50 (Tischer VZV-BAC [18]), as detailed in the text. In VZV-ORF57-Luc, ORF13 (TS), ORF36 (TK), or both, were deleted through site-directed recombination events to replace the partial (ORF36) or entire (ORF13) ORFs with either the mturq2blue (mT2B) fluorescent reporter gene or a kanamycin resistance cassette. The colors used for each virus are carried through the rest of the figures for continuity.