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. 2014 Aug 20;9(8):e105520. doi: 10.1371/journal.pone.0105520

Figure 1. Schematic of cassettes inserted into recombinant adenovirus and MVA vectors.

Figure 1

A) Bi-cistronic transgene expression cassette for insertion into E1 locus of HAdV5 vector. A human CMV immediate early promoter (CMV) drives expression of the mouse 4-1BBL open reading frame, and the human EF1α promoter (EF1a) drives expression of the TIP-EGFP fusion protein, used as a model antigen. Two different polyadenylation signals (pA) are used. This assembly is flanked by AttL sites for Gateway cloning (G) into the HAdV-5 vector genomic clone. Ruler is in nucleotides. B) Tri-cistronic transgene expression cassette for insertion into TK locus of MVA vector. The early-late vaccinia virus p7.5 promoter drives expression of the TIP model antigen (not fused to GFP) and the early-late short synthetic promoter (SSP) drives expression of mouse 4-1BBL, while the late fowlpox virus p4B promoter drives expression of the GFP marker gene. This assembly is flanked by sequences from the viral thymidine kinase locus (TKL and TKR) to allow recombination into the viral genome in infected cells. Ruler is in nucleotides.