FIG. 2.

Kinetics of transcription of UL132. (A) Total RNA was isolated from infected fibroblasts and RT-PCR was carried out using primers specific for UL132 and glyceraldehyde-3-phosphate dehydrogenase (GAPDH). Amplification products were separated on a 1% agarose gel and stained with ethidium bromide. For immediate-early RNA preparation (lane IE), protein synthesis was inhibited using cycloheximide and RNA was prepared 7 h after infection. For preparation of early RNA (lane E), phosphonoformic acid was used to block viral DNA synthesis and the RNA was prepared 24 h after infection. Late RNA (Lane L) was prepared 72 h postinfection. Lane PCR represents a control PCR without reverse transcription. (B) Northern blot analysis of total RNA isolated from fibroblasts infected with HCMV strain AD169 or TB40E. RNA was isolated at the indicated hours postinfection (hpi) and subjected to Northern blot analysis. A strand-specific probe from the UL132 ORF was used to detect the RNA. Detection of glyceraldehyde-3-phosphate dehydrogenase (GAPDH) RNA was used as a control.