Figure 2From: The duck hepatitis virus 5'-UTR possesses HCV-like IRES activity that is independent of eIF4F complex and modulated by downstream coding sequencesThe 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.Back to article page