Figure 1
Construction of chimeric rAd5 vectors containing hexon HVRs from Ad37 and Ad43.
Recombinants selected by GFP expression were identified by PCR and their recombination efficiencies were calculated. (a) A schematic strategy to construct Ad5-37(5, 7), Ad5-37(1–7), Ad5-43(5, 7) and Ad5-43(1–7) vectors by exchanging either the fifth and seventh HVR or all seven HVRs with the corresponding regions. The Ad5 genes with the hexon protein, indicated in blue, are shown above. HVR sequences from Ad37 are indicated in green and Ad43 in orange. (b) The HVR-chimeric Ad5 genome plasmids together with the shuttle plasmid expressing GFP were co-transfected into HEK293 cells (1 × 106 cells in 10 cm2 dish), single cytopathic effect (CPE) plaques expressing GFP were counted 10 days after co-transfection. (c) HEK293 cells were infected with the indicated virus at an MOI of 10 IFU/cell and CPE images were taken 48 h after infection with the indicated chimeric adenovirus. (d) HEK293 cells were infected with the indicated virus at an MOI of 10 IFU/cell, chimeric adenoviruses were harvested at the indicated time points from cells by three freeze-thaw cycles and vector yield was determined using rapid titer method. Data are means ± SD (n = 3).
