Fig. 4. Analysis of the DNA content and protein composition of purified wt and mutant particles.
A) Samples of purified wt and Cts52 were diluted and the amount of virions determined by A260nm and indicated on the top of each lane were applied onto a nylon membrane, processed and hybridized to 32P VACV DNA fragment. B) The blot was analyzed with a phosphor-imager apparatus, the data were normalized and graphed as described in Methods. C) The polypeptide composition of purified virions was analyzed by SDS-PAGE and Coomassie blue staining. The amount of virions determined by A260nm is indicated on the top of each lane and the approximate molecular weight marker is indicated on the left of the gel. D) Partition of purified virions into core and membrane fraction and Western blot analysis. 0.06 A260 nm units (~0.7 μg) of each virion sample was resuspended in core buffer and processed as described under Methods. The gene product corresponding to each antiserum used with the predicted molecular weight is indicated to the left of each row, the virus sample probed is indicated on the top of the figure. C= core fraction, M = membrane fraction. Wt31, wt grown at 31°C; Wt-40, wt grown at 40 °C; Cts- 31, Cts52 grown at 31°C; Cts40, Cts52 grown at 40 °C.