Figure 1 | Scientific Reports

Figure 1

From: Novel recombinant chimeric virus-like particle is immunogenic and protective against both enterovirus 71 and coxsackievirus A16 in mice

Figure 1

Schematic representation of the ChiEV-A71-P1 constructs and expression and purification of ChiEV-A71 VLP.

(A) The DNA sequences coding for the SP70 epitope in EV-A71-P1 was replaced with the corresponding region in CVA16-P1 to obtain ChiEV-A71-P1. The genes were under controls of GAL1 inducible promoters. (B) SDS-PAGE analysis indicated that fractions 11–16 contained enriched capsid proteins, i.e., VP0, VP1 and VP3, suggesting likely the presence of VLPs. The density of these fractions is about 1.13 g/ml. (C) Fractions were taken from top to bottom after sucrose gradient ultracentrifugation (10–40%). Samples of the fractions were probed with antibodies against VP0, indicating the expression of both ChiEV-A71-P1 and 3CD proteins and efficient protease activity of the expressed 3CD in the yeast cytoplasm. (D) SDS-PAGE analysis of purified ChiV71 VLPs. Fractions 11–16 were collected, diluted with PBS and concentrated by ultracentrifugation. Coomassie blue staining of the purified ChiEV-A71 VLPs demonstrated the presence of three capsid proteins: VP0 (38 kDa), VP1 (36 kDa) and VP3 (27 kDa).

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