VP22 fusion protein-based dominant negative mutant can inhibit hepatitis B virus replication.
OBJECTIVE
To investigate the inhibitory effect of VP22 fusion protein-based dominant negative (DN) mutant on Hepatitis Bvrus (HBV) replication.


METHODS
Full-length or truncated fragment of VP22 was fused to C terminal of HBV core protein (HBc), and subcloned into pcDNA3.1 (-) vector, yielding eukaryotic expression plasmids of DN mutant. After transfection into HepG2.2.15 cells, the expression of DN mutant was identified by immunofluorescence staining. The inhibitory effect of DN mutant on HBV replication was indexed as the supernatant HBsAg concentration determined by RIA and HBV-DNA content by fluorescent quantification-PCR (FQ-PCR).Meanwhile, metabolism of HepG2.2.15 cells was evaluated by MTT colorimetry.


RESULTS
VP22-based DN mutants and its truncated fragment were expressed in HepG2.2.15 cells, and had no toxic effect on host cells. DN mutants could inhibit HBV replication and the transduction ability of mutant-bearing protein had a stronger inhibitory effect on HBV replication. DN mutants with full length of VP22 had the strongest inhibitory effect on HBV replication, reducing the HBsAg concentration by 81.94%, and the HBV-DNA content by 72.30%. MTT assay suggested that there were no significant differences in cell metabolic activity between the groups.


CONCLUSIONS
VP22-based DN mutant can inhibit HBV replication effectively.
Yi
Jun
J
Department of general Surgery, Xijing Hospital, Fourth Military Medical University, Shaanxi Province, China.
Gong
Wei-Dong
WD
Wang
Ling
L
Ling
Rui
R
Chen
Jiang-Hao
JH
Yun
Jun
J
D006528:Carcinoma, Hepatocellular; D045744:Cell Line, Tumor; D015316:Genetic Therapy / Q000379:methods*; D006509:Hepatitis B / Q000628:therapy* / Q000821:virology; D006515:Hepatitis B virus / Q000235:genetics / Q000254:growth & development*; D006801:Humans; D008113:Liver Neoplasms; D016296:Mutagenesis; D014760:Viral Fusion Proteins / Q000235:genetics; D015678:Viral Structural Proteins / Q000235:genetics*; D014779:Virus Replication
D014760:Viral Fusion Proteins; D015678:Viral Structural Proteins; C073413:herpes simplex virus type 1 protein VP22