FIG. 5.

Expression of IE19 mRNA. (A) The MIE gene and its transcripts. RT-PCR primers and the predicted sizes of amplified cDNAs are also shown. (B) Expression of IE19, IE72, and UL112-113 mRNAs was examined by RT-PCR. HEL cells were infected with HCMV (Towne) at an MOI of 5 and harvested at 0, 3, 6, 12, 24, 48, and 72 hpi for RNA preparation. Total RNA (1 μg) was used for each RT-PCR. The PCR cycle was repeated 30 times. The products were examined by Southern blot hybridization with an oligonucleotide probe specific for IE72 and UL112-113 mRNAs. As an internal control, expression of β-actin mRNA was also examined by RT-PCR. An ethidium bromide staining image of the agarose gel is shown for β-actin. (C) HEL cells were cultured with cycloheximide (200 μg/ml) for 2 h before infection and maintained in the cycloheximide-containing medium after infection. Viral infection and analysis of total RNA were performed as described for panel A. The nucleotide sequences of these RT-PCR primers and probes are shown in Table 1.