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. 2023 Jun 21;15(7):1409. doi: 10.3390/v15071409

Figure 1.

Figure 1

Construction of recombinant LSDV-SARS2-S,N. (A) Schematic diagrams of the wild type and modified SARS-CoV-2 spike antigens with amino acid residue positions labeled above. The native signal sequence (SS) was replaced with the tissue plasminogen activator signal sequence (TPA), the furin cleavage site (RRAR) was replaced with a short linker sequence (GSAS), and two stabilizing proline mutations were included (K986P and V987P). (B) Schematic diagram of the transfer vector LSDV-Red SARS-CoV2-S+N used for homologous recombination with the parent virus LSDV(SODis)BEFV-Gb genome between ORFs 49 and 50 to generate LSDV-SARS2-S,N. (C) Generation and passage of LSDV-SARS2-S,N. The recombinant virus (mCherry; red) was constructed in LT cells at passage 0 (P0) by infection with LSDV(SODis)BEFV-Gb (eGFP; green) and transfection with the transfer vector. P1 = first passage in MDBK cells, P2 = second passage in MDBK cells, P3 = third passage in MDBK cells. All scale bars = 50 µm.