Expression of influenza A virus-derived peptides on a rotavirus VP6-based delivery platform

Figures

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  Fig. 1. Schematic representation of insertion sites on the rotavirus VP6 protein and fusion protein design. (A) VP6 homotrimer with different insertions sites at aa 171, aa 301 and aa 311 located in different surface loops, N-terminus (N-t), and C-terminus (C-t), which are indicated by arrows. The VP6 structure was modelled in SWISS-MODEL using the crystal structure solved by Mathieu et al. [18] as a template (PDB code: 1qhd). (B) Schematic representation of seven fusion proteins (FPs) carrying the M2e peptide and/or the HA stem fragment polypeptide from influenza A virus at the N- or C-terminus or internal sites of VP6. Flexible linkers were used to connect independent fragments.

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  Fig. 2. Expression of VP6 fusion proteins carrying influenza A virus-derived antigens. (A) Pellet (P) and supernatant (S) samples from fusion proteins FP1-FP7 immunoblotted with VP6-specific antibody. M (molecular weight)-marker in kilodaltons (kDa); wtVP6, purified "wild-type" VP6 protein (45 kDa) used as a positive immunoblotting control. The figure is composed of blots from several different experiments, which are separated by lines. All samples, however, were subjected to same procedures during the experiments. (B) Pellet samples of FP1, FP3, FP4 and FP7 and the supernatant sample of FP2 immunoblotted with M2-specific antibody. (C) The supernatant samples of FP5 and FP6 and the pellet sample of FP7 immunoblotted with HA-specific antibody