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. Author manuscript; available in PMC: 2021 Nov 1.
Published in final edited form as: Virology. 2020 Aug 22;550:51–60. doi: 10.1016/j.virol.2020.08.003

Figure 1. Generation of replication competent recombinant A/PR8 viruses containing chimeric HA constructs with a conserved epitope inserted into the head domain.

Figure 1.

Recombinant A/PR8 influenza viruses containing chimeric NAe-HA and M2e-HA were generated via the reverse genetics system using the attenuated pHW2000-based eight-plasmid system. (A) Chimeric HA constructs with a conserved epitope inserted into the head domain. NA222–230 (NAe-HA) or human M2e7–16 (M2e-HA) were inserted between positions 171 and 177 as indicated by the color boxes, respectively. (B) In vitro growth of recombinant A/PR8 influenza viruses containing chimeric HA. The 293T cell culture supernatants at 48h from co-transfected with eight pHW2000 plasmids including the chimeric HA constructs and attenuated PB1 and PB2 genes were inoculated into embryonated chicken eggs. Hemagglutination activity units (HAU, left panel) and the egg infective dose titers (right panel) in allantoic fluids containing rescued virus were determined using the chicken red blood cells and an egg infection assay. EID50; 50% egg infective dose. (C) Size distribution of influenza PR8 viruses containing chimeric HA molecules.