Figure 3.

Schematic representation of Kawagishi’s reverse genetics system. BHK-T7 cells are transfected with a set of 11 rescue T7 plasmids encoding an RVA genome as well as five helper expression plasmids for the NSP2, NSP5, VV-capping enzyme, and small membrane fusion proteins. After 2 days, the transfected BHK-T7 cells are overlaid with RVA-susceptible cells (MA104 cells) and then co-cultured for 5 days for amplification of recombinant RVAs. Recombinant RVAs can be rescued from the co-cultures. PT7, Rib, PCAG, and FAST denote the T7 RNA polymerase promoter sequence, HDV ribozyme sequence, CAG promoter sequence, and small membrane fusion protein, respectively.