FIG. 7.
Effect of N-terminal 2A deletions on 3Cpro-mediated processing of the capsid protein precursors expressed by recombinant VVs. (A) Schematic showing HAV polyprotein expressed by the recombinant VV containing the open reading frame, as well as the 3′ nontranslated region, of the v18f genome downstream of the EMCV internal ribosome entry site and the T7 RNA polymerase promoter (black triangle). Enlargements are shown for the 2A deletions introduced into the polyproteins of the indicated mutants. The positions within the HAV polyprotein of amino acids framing each deletion are indicated in boxes. FRhK-4 cells were infected with vTF7-3 (Mock) or coinfected with vTF7-3 and either the parent virus (VV-18f) or the indicated mutant virus (VV-Δ2A-1, -3, or -5), each at an MOI of 5 PFU/cell. HAV proteins from cytoplasmic extracts prepared at 20 hpi were separated by SDS-10% PAGE and identified by immunoblotting using anti-2B antibodies (B) or a mixture of anti-VP1 and anti-VP2 antibodies (C). HAV polypeptides, as well as molecular mass markers, are indicated on each side of the panels.