Construction of whole-genome lamivudine-resistant hepatitis B virus mutant and its replication and expression in HepG2 cells / 南方医科大学学报

ABSTRACT

<p><b>OBJECTIVE</b>To observe the changes of replication and antigen expressions of lamivudine-resistant hepatitis B virus (HBV) with polymerase gene mutation.</p><p><b>METHODS</b>With site-directed mutagenesis, we constructed 3 HBV plasmids with polymerase gene mutation based on the template P3.8II. All the plasmids were transfected into HepG2 cells, in which the replication and expression of the virus were analyzed.</p><p><b>RESULTS</b>The 3 polymerase gene mutant HBVs were successfully constructed. HBsAg and HBeAg expressions were detected in the supernatants of HepG2 cells transfected with these plasmids. The replication of the 3 mutant plasmids with rtG172E, rtG174C and rtG172E/rtG174C mutation decreased significantly in comparison with the wild-type virus.</p><p><b>CONCLUSION</b>The 3 polymerase gene mutant HBVs shows depressed capacity for viral replication, and in vitro drug sensitivity study is needed to establish the relationship between these mutations and nucleoside analogue resistance.</p>