FIG 1.

Expression of WT and mutant HBc proteins in RRL. (A) Schematic diagram of HBc domain structure and the CTD sequence. The three major phosphorylation sites in CTD (S155, S162, and S170) are marked above the sequence and the four minor sites (T160, S168, S176, and S178) below. pCI-HBc-3A (3A) and 3E have the three major sites changed to A and E, respectively, whereas pCI-HBc-7A (7A) and 7E have all seven sites substituted. (B) Estimation of HBc concentration as expressed in RRL. Each lane contained 3 μl of the translation reaction mixture. The ³⁵S-labeled HBc proteins were detected by autoradiography (top) and by Western blot analysis using the anti-HBc NTD MAb (bottom). (C) Translation in RRL and capsid assembly schemes. The five different assembly conditions are listed at the bottom. See the text for details. C, WT HBc protein; C149, C-terminally truncated HBc protein (terminated at position 149); CIAP, calf intestine alkaline phosphatase; PPI, phosphatase inhibitors.