4.4. Plasmids, Transfections, and Luciferase Reporter Assays

HCMV MIE enhancer/promoter sequences were removed from the vector pJHA324 using the restriction enzyme HindIII (Enzynomics, Daejeon, Korea) [33]. The VZV IE promoter was synthesized (Macrogen, Seoul, Korea) and amplified by PCR using a primer containing the HindIII sequence at the end. The primer sequences used for amplification were 5′-CCC AAG CTT ATC GTC TGT AGA CAC ACG ATG-3′ (forward) and 5′-CCC AAG CTT CGC ACT GGG GTG AAT TTA G-3′ (reverse). The PCR product was digested with HindIII and ligated using T4 ligase (Enzynomics) into a pJHA324 vector in which the HCMV MIE enhancer/promoter sequences had been removed. The orientation of the VZV IE promoter insert was confirmed by DNA sequencing (Macrogen, Seoul, Korea). Transient transfections and luciferase assays were performed using Omicsfect™ (Omics Bio, Taipei city, Taiwan) and Dual-Luciferase Reporter Assay System (Promega, Madison, WI, USA), respectively, according to the manufacturers’ protocols as described previously [34].