FIG. 3.

Detection of ORF-E in TG of BHV-1-infected calves. Calves were infected with BHV-1 as previously described (14, 30). At 4, 6, 8, 14, and 60 days postinfection, TG were collected and total RNA was extracted as previously described (6, 13). (A, B, and C) Strand-specific RT-PCR was performed to detect expression of ORF-E RNA with primer 308. ORF-E-specific transcripts were detected by PCR with primers 478 and 598. Panels A and B show a representation of two different experiments. The number of days that the calves were infected is denoted on the top of the panels. L, RNA extracted from TG of latently infected calves (60 days after infection), included to give a relative level of ORF-E RNA detection between acutely and latently infected tissue. (D and F) Random-primed RT-PCR was performed to detect gC and β-actin RNA expression, respectively. (E) Strand-specific RT-PCR was performed to detect expression of the LR RNA with the L3B reverse primer, and PCR was performed with the L3B primers. The arrow denotes the expected PCR product. The bands below the expected products are primer dimers. Lanes: M, mock-infected calf; L, latently infected calves; +, DNA from BHV-1-infected MDBK cells; −, no-template control.