Expression of BHRF1 and Northern Blot

Expression of BHRF1 mRNA was analyzed by an RT-PCR based assay as previous described [150]. In brief, 1 μg of RNA was reversed transcribed and used to perform PCR reaction with BHRF1 specific primers (5'-GTC AAG GTT TCG TCT GTG TG-3' and 5'-TTC TCT TGC TGC TAG CTC CA-3') and actin specific primers (5'-TAA GGA GAA GCT GTG CTA CGT C-3' and 5'-GGA GTT GAA GGT AGT TTC GTG G-3). PCR products were analyzed in 2% agarose gel and transferred onto Nytran SPC membrane (Whatman Inc, Piscataway, NJ) for Southern blot analysis using γ-32P ATP end-labeled synthetic oligonucleotides complementary to the lytic form of BHRF1 (5'-ATG CAC ACG ACT GTC CCG TAT ACA C-3'). Northern blot was performed as described previously [23,122]. The sequence of synthetic oligonucleotides used as probes for EBER1-5p and 3p were EBER1-5p: 5'-TAG GGC AGC GTA GGT CCT-3' and EBER1-3p: 5'-AAA CAT GCG GAC CAC CAG CTG G-3'. As a positive control, 5 μg of RNA from EBV-positive marmoset cells line B95-8 was included for Northern blot analysis.
