The DHV 5'UTR initiates translation both in vitro and in vivo. (A) Dicistronic capped RNAs were in vitro generated from plasmid containing a cDNA insert corresponding to the DHV-1 5'-UTR (DHV UTR), pGEM-CAT/EMCV/LUC (EMCV-IRES) or pGEM-CAT/LUC (CAT/LUC). RNAs were individually added to Flexi rabbit reticulocyte lysate (RRL) system (Promega) and the in vitro translation of the reporter proteins was evaluated measuring 35S-methionine incorporation. (B) The dicistronic plasmids were transfected into vFT7-infected BHK-21 cells, and 24 h post-transfection, cell extracts were prepared and protein synthesis was analyzed by SDS-PAGE and immunoblotting to detect CAT and fLUC expression.
Liu et al. Virology Journal 2011 8:147 doi:10.1186/1743-422X-8-147