Study of IFN-inducible double-stranded RNA dependent protein kinase on antiviral activity of HBV in vitro
10.3969/j.issn.1001-1978.2015.09.015
- VernacularTitle:干扰素诱导的双链 RNA 依赖性蛋白激酶体外抗乙型肝炎病毒活性的研究
- Author:
Aihua WANG
;
Shihe GUAN
;
Kai YANG
;
Hao ZHANG
;
Beibei SUN
;
Ying PAN
;
Jilong SHEN
- Publication Type:Journal Article
- Keywords:
hepatitis B virus;
plasmid;
PKR protein;
antigen;
complication;
cells
- From:
Chinese Pharmacological Bulletin
2015;(9):1254-1258
- CountryChina
- Language:Chinese
-
Abstract:
Aim To construct and express the eukary-otic expression vector of double-stranded RNA-depend-ent protein kinase (PKR)fusion green fluorescent and analyse its antiviral activity of HBV in vitro.Methods The PKR gene was cloned into an empty expression vector pEGFP-N1 using molecular clone technology. After being confirmed by restriction enzyme digestion and sequencing methods,the recombinant plasmid was named as pEGFP-PKR that was subsequently transfect-ed into HepG2.2.15 cells using LipofectamineTM2000. The expression level of PKR in HepG2.2.15 cells was confirmed by using fluorescent microscopy. Mean-while,HBV DNA and HBsAg/HBeAg were detected by real-time PCR and electrochemiluminescence meth-od,respectively.Results Both restriction enyme di-gestion and sequencing assays showed that the recombi-nant vector pEGFP-PKR was successfully constructed in our study.Fluorescent microscopy observation indi-cated that the fusion protein pEGFP-PKR expressed ef-ficiently in HepG2.2.15 cells.Moreover,compared with the empty vector group,the expression of HBV antigen in supernatants was significantly decreased (P<0.05 ).However,the extracellular HBV DNA ex-pression was not inhibited significantly.Conclusion In vitro,PKR proteion has certain antiviral activity of HBV.
