FIG 5.

Analysis of capsid assembly in RRL by sucrose gradient centrifugation. The indicated RRL translation reaction mixtures (100 μl), either left untreated or treated as indicated, were separated over a linear 15% to 30% sucrose gradient spun in an SW55 rotor at 27,000 rpm for 4 h at 4°C. The indicated sucrose fractions (10 μl each) or input RRL translation mixtures (either left untreated or treated as indicated) (0.5 μl) were resolved by agarose gel electrophoresis. The direction of centrifugation is indicated by the arrows. ³⁵S-labeled HBc proteins were detected by autoradiography. Capsids purified from E. coli (unlabeled) in panel C were detected by Western blot analysis using the anti-HBc NTD MAb. Fract #, sucrose fraction number; mock, overnight incubation in the assembly buffer alone; C and 3E, full-length HBc proteins; C149, C-terminally truncated HBc protein (terminated at position 149); Ca, capsids.