Preparation and identification of a recombinant hepatitis C virus (HCV) based on sindbis virus vector.
- Author:
Juan-Juan FU
1
;
Wu-Yang ZHU
;
Jiang-Jiao LI
;
Huan-Qin WANG
;
Wen-Jie TAN
;
Jian-Zhong YIN
;
Guo-Dong LIANG
Author Information
- Publication Type:Journal Article
- MeSH: Animals; Cells, Cultured; Cricetinae; Genetic Vectors; Hepacivirus; genetics; Plasmids; Recombination, Genetic; Sindbis Virus; genetics; Viral Envelope Proteins; genetics
- From: Chinese Journal of Experimental and Clinical Virology 2012;26(2):136-138
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVETo construct the recombinant virus-like particles containing HCV envelope glycoprotein E1E2 based on sindbis virus vector.
METHODSThe gene encoding HCV envelope glycoprotein E1E2 was cloned into sindbis virus vector to construct recombinant plasmids pBR-XJE1E2 and pVA-XJE1E2, and transfect them into BHK-21 cells to obtain recombinant virus-like particles. The expression of E1 and E2 protein were verified by Western Blot and indirect immunofluorescent assay (IFA).
RESULTSThe results of restriction enzyme digestion, PCR and sequencing analysis showed that the recombinant plasmids were constructed successfully. And the results of RT-PCR, Western blotting and IFA detection showed that the transfect cells could package HCV-like particles of expressing structural proteins E1E2.
CONCLUSIONThe recombinant expression plasmids pBR-XJE1E2 and pVA-XJE1E2 based on sindbis virus vector could package HCV-like particles in eukaryotic cell, which provides a foundation for further study of its in vivo animal immune response.
