Effect of viral gene deletion and mutation in the UL133–138b region on recombinant TR viruses.

<p>HFF cells were infected with recombinant wt and mutant TR virus at an MOI of 0.5 PFU/cell. Separate batches of cells were harvested for both protein and DNA analysis. (A) Western blot analysis to determine equivalent virus particle input by quantifying the pp65 protein associated with the infected cells at 3 h p.i. (B) Western blot analysis of MIE and late viral protein. GAPDH served as a loading control. (C) Quantification of gB DNA normalized to 18S DNA using real time PCR. The insert shows equal viral DNA input at 3 h p.i. (D) Total virus at various d p.i. Cells and supernatant were harvested and subjected three times to freeze and thaw lyses. (E) Extracellular virus at various d p.i.