[Construction of whole-genome lamivudine-resistant hepatitis B virus mutant and its replication and expression in HepG2 cells].
OBJECTIVE
To observe the changes of replication and antigen expressions of lamivudine-resistant hepatitis B virus (HBV) with polymerase gene mutation.


METHODS
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.


RESULTS
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.


CONCLUSIONS
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.
Sun
Jian
J
Department of Infectious Diseases, Nanfang Hospital, Southern Medical University, Gunagzhou 510515, China. sunjian@fimmu.com
Wang
Cheng
C
Wen
Shu-juan
SJ
Wang
Zhan-hui
ZH
Hou
Jin-lin
JL
D001483:Base Sequence; D024882:Drug Resistance, Viral / Q000235:genetics*; D015870:Gene Expression; D005818:Genetic Engineering / Q000379:methods*; D016679:Genome, Viral / Q000235:genetics*; D056945:Hep G2 Cells; D006514:Hepatitis B Surface Antigens / Q000235:genetics; D006513:Hepatitis B e Antigens / Q000235:genetics; D006515:Hepatitis B virus / Q000187:drug effects / Q000235:genetics* / Q000276:immunology / Q000502:physiology; D006801:Humans; D019259:Lamivudine / Q000494:pharmacology*; D009154:Mutation; D010957:Plasmids / Q000235:genetics; D014162:Transfection; D014779:Virus Replication / Q000235:genetics*
D006514:Hepatitis B Surface Antigens; D006513:Hepatitis B e Antigens; D019259:Lamivudine