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. 2015 Sep 23;89(23):12221–12225. doi: 10.1128/JVI.02382-15

FIG 1.

FIG 1

Construction and growth of a UL93 stop mutant virus. (A) Schematic of the UL92-to-UL94 region in TB40/E-BAC (top line) with other bacmid constructs shown below. A UL93 stop mutant virus (UL93st-TB40/E-BAC) was constructed using two-step BAC recombineering (17). In the first step, a kanamycin (Kan) cassette fused to the UL93 stop mutation and an I-SceI site was inserted in the BAC genome using homologous recombination. The Kan cassette was then spliced out utilizing the I-SceI site, leaving only the UL93 stop mutation in the genome. (B) Final virus yields were determined at day 10 postinfection for wild-type TB40/E (WT-TB40/E in HF), UL93st-TB40/E-BAC in complementing cells (UL93st in FLAG-UL93 HF), and UL93st-TB40/E-BAC in noncomplementing cells (UL93st in HF). N.D., not detected, as the number of PFU in this preparation was below the threshold of detection in the assay.