FIG. 4.

ADdlUL38 fails to replicate viral genomes to wild-type levels. (A) Accumulation of viral DNA. Fibroblasts were infected with ADdlUL38 or ADwt at an input genome number equivalent to 0.2 PFU of wild-type virus/cell, total cell-associated DNA was isolated, and viral genomes were quantified using real-time PCR and normalized to β-actin DNA. (B) Accumulation of the UL123 transcript and the protein product IE1. Fibroblasts were infected under the conditions described above. Total RNA was isolated at the indicated times postinfection, and UL123 RNA was quantified by real-time RT-PCR using primers specific to UL123, with SYBR green detection, and normalized to GAPDH RNA. Accumulation of the IE1 protein was determined by Western blot analysis. An antibody to alpha-tubulin was used as a loading control. (C) Accumulation of the UL99 transcript and the protein product pp28. The UL99 transcript was measured by real-time RT-PCR using primers specific to UL99. The pp28 protein was detected by Western blot analysis using an antibody to pp28.