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

Figure 2.

Figure 2

Schematic representation of Sánchez-Tacuba’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, as well as a chimeric helper expression plasmid for the African swine fever virus-capping enzyme NP868R and T7 RNA polymerase. Next day, the transfected BHK-T7 cells are overlaid with genetically modified MA104N*V cells with impaired innate immune responses and then co-cultured for 3 days for amplification of recombinant RVAs. Infectious recombinant RVAs can be rescued from the co-cultures. PT7, Rib, and PCMV denote the T7 RNA polymerase promoter, HDV ribozyme, and cytomegalovirus promoter sequence, respectively.