FIG. 5.
Reversion of the reduced viral DNA synthesis phenotype associated with the YVDD variant. HepG2 cells were transfected with 4 μg of pCMVhbv (YMDD), pAT0304 (YVDD), or pAT0307 (YMDDR). Three days after transfection, immature core particles were isolated as previously described (6). Viral DNA was released from the core particles by proteinase K treatment followed by phenol extraction and ethanol precipitation. It was then separated by size by electrophoresis, transferred to a nylon filter, and detected with a radioactively labeled HBV-specific probe. The autoradiography exposure time was 16 h. Note that there was a 10-fold difference in the amount of viral DNA produced by pCMVhbv and pAT0307 and pAT0304 after normalization to transfection efficiency. rc, relaxed circular; ln, linear; ss, single stranded.