FIG. 2.

Biochemical and electron microscopy analyses of the structures generated by expression of IBDA-GFP polyproteins modified at the C terminus of pVP2. (A) After an overnight isopycnic centrifugation, each gradient corresponding to one of the four deletion mutants was illuminated with a white (left) or UV (right) light and photographed. A band is visible for mutants Δpep46, Δpep7a, and Δpep7b. The bands associated with Δpep7a and Δpep7b are fluorescent, in contrast to the Δpep46 band. Samples collected from the bands were concentrated and negatively stained with 2% uranyl acetate solution. (B) Material extracted from each band shown in panel A was analyzed by SDS-PAGE followed by Coomassie blue staining (left) or Western blot analyses with anti-pVP2/VP2 or anti-VP3 monoclonal antibodies (right). (C) Gradients corresponding to substitution mutants in the pep11 domain were photographed. Concentrated samples were processed for electron microscopy analysis. (D) Material extracted from each band shown in panel C was analyzed by SDS-PAGE followed by Western blot analyses with anti-pVP2/VP2 or anti-VP3 monoclonal antibodies.