FIG. 1.

Examination of BHV-1 transcripts during reactivation from latency. (A) The primers shown were previously used to detect viral transcripts in TG of infected calves (37). (B) RNA was prepared from MDBK cells infected with the LR mutant virus or the LR rescued virus (6 h after infection). cDNA synthesis was performed as previously described (37). RT reactions with (+) or without (−) reverse transcriptase are shown. DNA from BHV-1- and mock-infected MDBK cells were used as positive (C⁺) and negative (C⁻) controls, respectively, for the PCRs. The first lane contains a 100-bp ladder (NE Biolabs). Primers used for these studies are shown in panel A. (C) To initiate reactivation from latency, calves latently infected (60 days after infection) with the LR rescued virus or the LR mutant virus (LR−) were given a single intravenous injection (jugular vein) of DEX (100 mg). At 24 or 48 h after DEX treatment, total RNA was prepared from each TG as previously described (14, 37, 45-47). cDNA synthesis was primed using poly(dT), and then PCR was performed with the designated primers listed in panel A. In a separate cDNA synthesis reaction, the bICP0 primer was used to examine bICP0 expression. The results are summarized in the graphs from TG of three calves/time point. Each ganglion was treated separately.