[Zinc finger protein designed to hepatitis B virus core promoter inhibit  the transcription of HBV in HepG2.2.15 cells].
OBJECTIVE
To observe the inhibition of hepatitis b virus(HBV) transcription in vitro by zinc fingrer protein(ZFP) which was designed specifically to bind DNA sequence in the HBV core promoter(Cp).


METHODS
The sequence of HBV Cp was submitted to the Zinc Finger Tools and selected a sequence from the Cp as the target site of ZFP. The nucleic acid sequence of ZFP was synthesised and cloned into pEGFP-N1 or pcDNA3.1(+) to construct the recombinant plasmid pEGFP-N1/ZFP-Flag or pcDNA3.1(+)/ZFP. COS-7 cells were transfected with pEGFP-N1 or pEGFP-N1/ZFP-Flag, the expression of ZFP was observed by green fluorescent microscope, RT-PCR and Western blot. HepG2.2.15 cells were transfected with pcDNA3.1(+) or pcDNA3.1(+)/ZFP, HBeAg and HBV DNA levels in cell supernatant were detected by ELISA and FQ-PCR, HBV mRNA was tested by RT-PCR.


RESULTS
ZFP can express in COS-7 cells.In th presence of ZFP, HBeAg expression and HBV DNA level was significantly reduce(P<0.05), and HBV mRNA was dramatically decreased.


CONCLUSIONS
ZFP can express in eukaryotic cells and inhibit the transcription of HBV in vitro.
Department of Hepatobiliary Surgery, Chongqing Medical University, Chongqing, China. cong922@126.com
Cong
C
Chen
Xin
X
Li
Ning
N
Guo
Hua
H
Zhang
Zhong-jun
ZJ
Wu
D004261:DNA Replication; D004279:DNA, Viral; D015967:Gene Expression Regulation, Viral; D056945:Hep G2 Cells; D006513:Hepatitis B e Antigens / Q000235:genetics; D006515:Hepatitis B virus / Q000235:genetics*; D006801:Humans; D011401:Promoter Regions, Genetic; D014158:Transcription, Genetic; D016335:Zinc Fingers / Q000502:physiology*
D004279:DNA, Viral; D006513:Hepatitis B e Antigens