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. 2021 Sep 8;13(9):1791. doi: 10.3390/v13091791

Figure 1.

Figure 1

Schematic representation of the 11 plasmid-only reverse genetics system (Komoto’s reverse genetics system). BHK-T7 cells are transfected with a set of 11 rescue T7 plasmids encoding an RVA genome with 3-fold increased proportions of T7 plasmids for the NSP2 and NSP5 genes compared to the other nine T7 plasmids. Next day, the transfected BHK-T7 cells are overlaid with RVA-susceptible cells (CV-1 or MA104 cells) and then co-cultured for 3 days for amplification of recombinant RVAs. Infectious recombinant RVAs can be recovered from the co-cultures. PT7 and Rib denote the T7 RNA polymerase promoter and HDV ribozyme sequence, respectively.