Predicted domain IIIe and stem1 on DHV-1 5'-UTR are required for in vitro IRES activity. (A) The position of the different mutations evaluated in Figures 7 and 8 is shown in red. (B) To evaluate the relevance of the predicted structures in DHV-IRES, a dicistronic plasmid, in which the most important loop sequence within the putative IIIe region (GAUA) is mutated to AAAA (DHV IIIe mut), was constructed. A different dicistronic plasmid containing mutations to disrupt the predicted base pairing of stem 1 (DHV stem1 mut) and the corresponding reverse mutant that restores the structure (DHV stem1 mut') were also constructed. Mutant constructs were in vitro transcribed analyzed in RRL and translation of the reporter proteins was evaluated.
Liu et al. Virology Journal 2011 8:147 doi:10.1186/1743-422X-8-147