The effects of different PAP domains on hepatitis B virus replication.
- Author:
Chun-xia GUO
1
;
Yong-wen HE
;
Cheng PENG
;
Yan-chang LEI
;
Wen-ting LI
Author Information
- Publication Type:Journal Article
- MeSH: Amino Acid Sequence; Antiviral Agents; pharmacology; Blotting, Western; DNA, Viral; drug effects; metabolism; Hep G2 Cells; Hepatitis B Surface Antigens; metabolism; Hepatitis B e Antigens; metabolism; Hepatitis B virus; drug effects; genetics; physiology; Humans; Liposomes; Plasmids; genetics; Reverse Transcriptase Polymerase Chain Reaction; Ribosome Inactivating Proteins, Type 1; genetics; metabolism; pharmacology; Sequence Deletion; Transfection; Virus Replication; drug effects
- From: Chinese Journal of Hepatology 2010;18(2):105-108
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVETo investigate the effects of different PAP domains on hepatitis B virus replication.
METHODSThe full length and two truncated PAP mutants were cloned into a eukaryotic expression plasmid, and were transfected into HepG2.2.15 cells using lipofectamine 2000. 3 days after transfection, the medium and cells were collected. HBsAg and HBeAg were measured using ELISA. The titers of HBV DNA were quantified using fluorogenic quantitative PCR (FQ-PCR). HepG2 cells were used to determine the cytotoxicity of the plasmids transfection by MTT assays.
RESULTSThe inhibitory effect on HBV replication of the C-terminal 25 amino acids deleted PAP mutant (pXF3H-PAP14) was not significantly different from that of the full length PAP (pXF3H-PAP12) (Chi-square test = 0.5, 2.0, 0.02, probability value more than 0.05), however, the cytotoxicity of pXF3H-PAP14 was lower than that of pXF3H-PAP12 (Chi-square test = 7.7, probability value less than 0.01). Both N-terminal 69 amino acids deleted mutant and C-terminal 25 amino acids deleted mutant had no cytotoxicity and no antiviral activity.
CONCLUSIONC-terminal 25 amino acid of PAP is related to cytotoxicity but not related to antiviral activity of PAP. N-terminal 69 amino acid of PAP is related to the anti-HBV effect of PAP.
