Recombinant Protein Expression and Purification

The ABD gene was cloned into the previously prepared pET28a-VP1 plasmid through BamH I digestion sites. For evaluating dLNs accumulation of ABD-VP1 or VP1, the mCherry gene was cloned into the pET28a-ABD-VP1 or pET28a-VP1 plasmid. The inserted fragments were verified by DNA sequencing (Genewiz). Recombinant proteins were produced by the E.coli prokaryotic expression system and purified with His-beads. The specific expression and purification of those recombinant proteins were conducted as previously reported (22 (link)). Purified proteins were confirmed by western blot assays with the anti-VP1 antibody (Dako).