FIG 1 .

BADinUL71STOP generates fewer infectious progeny than BADwt. (A) Schematic of the BADinUL71STOP genome. An 11-base-pair translation stop cassette was inserted after nucleotide 36 of the UL71 ORF. (B) BADinUL71STOP produces >100-fold-fewer infectious progeny than BADwt. Infected fibroblasts and supernatants were harvested at the indicated times after infection at a multiplicity of 2 PFU/cell, and virus titers were determined by 50% tissue culture infective dose (TCID₅₀) assay. Inp, input virus. (C) Accumulation of RNA from neighboring genes is unaffected by the UL71STOP mutation. Fibroblasts were infected at a multiplicity of 2 PFU/cell with BADwt or BADinUL71STOP and harvested 72 h later. Total RNA was isolated. UL70 and UL72 message levels were quantified by reverse transcription-quantitative PCR (RT-qPCR) and normalized to actin RNA. Error bars represent standard errors of means calculated from technical repeats.