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. 2016 Jul 20;8(7):197. doi: 10.3390/v8070197

Figure 4.

Figure 4

Reporter-expressing recombinant arenaviruses. (A,B) R3 arenaviruses: For the generation of r3 arenavirus, the pPol-I plasmid expressing the S vRNA segment is separated in two S plasmids. In the pPol-I S1 plasmid, the viral NP is replaced by RG 1 and in the pPol-I S2 plasmid, the viral GPC is replaced by RG 2 (A); alternatively, RG 1 can be expressed instead of the viral GPC in the pPol-I S2 plasmid and RG 2 from the pPol-I S1 plasmid instead of the viral NP (B). Regulation of RG expression depends on their location in the S segment. Expression of an RG in the NP locus is higher than that observed when the RG is located in the GPC locus. The physical separation of the GP and NP proteins into two different S segments (S1 and S2) represents a strong selective pressure to maintain a virus capable of packaging one L segment and two S segments; (C) recombinant bicistronic arenaviruses: In the bicistronic rLCMV/GFP-P2A-NP, the NP open reading frame in the pPol-I S plasmid is replaced by the GFP-P2A-NP sequence that contains GFP tagged to the N terminus of NP, separated by the PTV1 2A peptide sequence (P2A). The P2A sequence allows for production of both GFP and NP from the same bicistronic mRNA transcribed from the NP locus of the S genome segment. Untranslated regions (UTR, black boxes) and intergenic regions (IGR) in each of the vRNA L and S segments are indicated.