Predicted domain IIIe, stem1 and stem2 on DHV-1 5'-UTR are required for in vivo IRES activity. A dicistronic plasmid containing mutations to disrupt the predicted base pairing of stem 2 (DHV stem2 mut) and the corresponding reverse mutant that restores the structure (DHV stem2 mut') were constructed. In vivo translation efficiency of these new plasmids was evaluated, together with the plasmids depicted in Figure 7. Briefly, dicistronic plasmids were transfected into DF-1 cells and 48 hpt the accumulation of fLUC and CAT protein expressed from the different constructs was evaluated. fLUC expression was normalized by CAT expression levels.
Liu et al. Virology Journal 2011 8:147 doi:10.1186/1743-422X-8-147