Figure 2. Mutation of VP2 internal methionine residues has little effect on MCV infectivity.

(A) Purified native MCV carrying mutations in the first internal methionine (Met₄₆) of VP2 (dVP3a), the second internal methionine (Met₁₂₉) of VP2 (dVP3b) or a double Met mutant (dVP3d) were analyzed alongside WT MCV by SDS-PAGE and western blot with a mixture of anti-VP1 and anti-VP2 rabbit sera. These native virus stocks were not produced simultaneously and differed slightly in concentration. Four µl of WT and dVP3d and 2 µl of dVP3a and dVP3b were run on the gel. (B) Equal amounts of WT and dVP3 mutant viruses, normalized by MCV genomic copies, were inoculated onto 293-4T cells. Samples taken one day or four days post-infection were analyzed for viral DNA concentration by qPCR to determine replication of virus-delivered genomic DNA as a measure of infectivity. The average of three experiments performed in duplicate is shown and error bars represent the standard error of the mean.