Figure 1

Schematic diagram of constructs and expression of vaccine plasmids. A. A diagram of the WNV genome is shown in the center, and the segments of the genome used in the vaccine constructs are shown above (DIII-modified) and below (prM/E-modified). The construct expressing prM/E was previously described [26]. The DIII region of the E gene (amino acids 586-705) was cloned downstream of the tpA leader sequence, and in some cases, P28 was also cloned in frame and directly after the 3' end of the DIII gene. An artificial BamHI site and stop codon was engineered at position 705 in the E gene to create the truncated Ecto E gene, and P28 was cloned into the Ecto E construct using the BamHI site to create the Ecto E-P28 construct. B. Supernatants from 293T cells transiently transfected with plasmid DNA were assessed by SDS-PAGE and Western blot. The membrane was probed by with the DIII-specific monoclonal antibody, 7H2. Lane 1: DIII-DNA; Lane 2: DIII-P28-DNA; Lane 3: prM/Ecto E-DNA; Lane 4: prM/Ecto E-P28-DNA; Lane 5: prM/E-DNA; Lane 6: P28-DNA only; Lane 7: Vector DNA only.