[Adenovirus-mediated transduction of HBV/C genome into HepG2 cell line].
OBJECTIVE
To establish a method for efficient transfer of 1.3-fold HBV/C genome into HepG2 cell line using adenoviral vector system for studying the replication and antigen expression of HBV.


METHODS
The 1.3-copy overlength genome of HBV genotype C was constructed and cloned into the shuttle vector pAdTrack. After confirmation of the constructed HBV genome by sequencing, the resultant plasmid linearized by digestion with Pme I was transformed into competent E.coli Adeasier-1 cells. The recombinants of pAdEasy-HBV/C were linearized by digestion with Pac I and transfected into the packaging cells (293 cells) via liposome. HepG2 cells were then infected with a proper quantity of the recombinant adenoviruses. The HBV DNA level and HBeAg and HBsAg titers were detected in the cell medium.


RESULTS
The 1.3-fold overlength HBV/C genome was efficiently transferred into HepG2 cells via the adenoviral vector system, which resulted in initiation of the virus replication and protein expression in the cells using the viral replication mechanism.


CONCLUSIONS
The adenovirus vector system (AdEasy) allows convenient and effective transfer of HBV genome into HepG2 cells, and provides a convenient means for screening therapeutic drugs against HBV.
Zhou
Bin
B
Department of Infectious Diseases, Nanfang Hospital, Southern Medical University, Guangzhou 510515, China. ice1126@163.com
Liang
Min-feng
MF
Li
Wei-dong
WD
Wang
Cheng
C
Wang
Zhan-hui
ZH
Hou
Jin-lin
JL
D000256:Adenoviridae / Q000235:genetics* / Q000378:metabolism; D006528:Carcinoma, Hepatocellular / Q000235:genetics / Q000821:virology; D005822:Genetic Vectors / Q000235:genetics; D016679:Genome, Viral / Q000235:genetics*; D006515:Hepatitis B virus / Q000235:genetics*; D006801:Humans; D008113:Liver Neoplasms / Q000235:genetics* / Q000821:virology; D014161:Transduction, Genetic; D014407:Tumor Cells, Cultured