Figure 1

The rAds constructed for this study. (A) Schematic representation of the linear genomes of the rAd-C and rAd-Bg viruses. The dashed line, close to the left end represents the position of region E1. The open box located towards the right end represents a ~2.7 Kb deletion in the non-essential E3 region (ΔE3). The hatched boxes at either end represent the left (L) and right (R) inverted terminal repeats (ITRs), and the open circle close to the L-ITR represents the packaging signal (ψ). In the rAds, region E1 is replaced either by an empty expression cassette (rAd-C) or the EDIII-4/2 expression cassette (rAd-Bg). Components of the expression cassettes are indicated by the following abbreviations, P: CMV promoter (the arrow indicates its orientation); 4 and 2: DEN-4 and DEN-2 EDIII-encoding regions, respectively, of the bivalent EDIII-4/2 gene; g: enhanced green fluorescent protein gene (fused in-frame to the EDIII-4/2 gene) and pA: SV40 polyadenylation signal. (B) Characterization of the rAd genomes by PCR. The picture depicts agarose gel analysis of PCR products generated using wt Ad5 (lanes 2, 6), rAd-C (lanes 3, 7) or rAd-Bg (lanes 4, 8) viral DNA templates. Primers used were either E1-specific (lanes 2–4) or specific to insert-flanking sequences (lanes 6–8). DNA size markers were run in lanes 1 and 5. Sizes of the lane 1 markers (in Kb) are indicated to the left of the panel. The position of the 1.6 Kb band in lane 5 is indicated by a white dot for orientation. The arrows to the right indicate the positions of the amplicons corresponding to the bivalent insert (top), E1 region (middle) and the insert-less vector sequences (bottom).