[Inhibitory effect of a HBV vector plasmid expressing A3C on HBV replication].
OBJECTIVE
To observe the inhibitory effect of a replication-defective hepatitis B virus (HBV) vector plasmid expressing A3C on HBV replication in vitro.


METHODS
The HBV vector plasmisd pCH-LJ3-A3C and pCH-LJ3-hrGFP expressing A3C and hrGFP were constructed using PCR and gene recombination technique. The two recombinant plasmids were separately cotranfected into HepG2 cells along with the wild-type HBV plasmid pCH-3093. The HBV DNA in the cell cytoplasmic lysates and in the cell culture supernatant was extracted for Southern blotting, and the nucleocapsid-associated HBV DNA were amplified by PCR, cloned and sequenced.


RESULTS
pCH-LJ3-A3C showed obvious inhibitory effect on HBV DNA in the cytoplasmic lysates and cell culture supernatant, causing a reduction of the HBV DNA by 31% and 40%, respectively. The pCH-LJ3-A3C plasmid was capable of editing the HBV DNA. Among the 50 sequenced clones, 36 clones had G-A mutations, with a total of 982 such mutations.


CONCLUSIONS
pCH-LJ3-A3C can inhibit the replication of HBV primarily by editing HBV DNA. The pCH-LJ3-A3C plasmid may serve as a new antiviral agent against human HBV infection.
Institute of Infectious Diseases, Southwest Hospital, Third Military Medical University, Chongqing 400038, China. lidong68@live.cn
Dong
D
Li
Yu-ming
YM
Wang
D000071480:APOBEC-3G Deaminase; D000998:Antiviral Agents / Q000494:pharmacology*; D001483:Base Sequence; D003564:Cytidine Deaminase / Q000235:genetics / Q000378:metabolism*; D005822:Genetic Vectors / Q000235:genetics; D056945:Hep G2 Cells; D006515:Hepatitis B virus / Q000235:genetics* / Q000502:physiology*; D006801:Humans; D008969:Molecular Sequence Data; D010957:Plasmids / Q000235:genetics / Q000378:metabolism; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D014779:Virus Replication / Q000235:genetics*
D000998:Antiviral Agents; D012333:RNA, Messenger; D000071480:APOBEC-3G Deaminase; C464924:APOBEC3G protein, human; D003564:Cytidine Deaminase