Figure 2

Schematic representation of norovirus protein coding region of pRN16 constructs expressing wild-type and hybrid VLPs. Following construction of plasmids pRN-GII4v0 and pRN-GII4v2 expressing wild-type GII-4v0 and GII-4v2 VLPs, respectively, these plasmid constructs were modified by site directed mutagenesis at either putative epitope site A (nt 886-891, aa 296-298), or putative epitope site B (nt 1176-1182, aa 383-395) to generate plasmid constructs that expressed hybrid VLPs. The schematic shows a representation of the region of the plasmid encoding norovirus structural proteins (3'UTR and remainder of plasmid not shown for clarity). PCR mutagenesis was used to generate plasmid constructs that encoded an ORF2 identical to either GII-4v0 or GII-4v2, except at either site A or site B, which was modified to be as equivalent to that position in the heterologous variant. The resulting expressed VP1 protein was a hybrid of the two variants, and so the VLP formed from the hybrid VP1 was antigenically hybrid. Plasmid construct names are given on the left, whilst the hybrid VLPs are represented with VLP names next to them on the right. All GII-4v0 derived regions are shown in solid black, all GII-4v2 derived regions are shown in hatched lines.