Development and Identification of the Recombinant Lentivirus Co-expressing HCV Structural Protein and Secreted Gaussia Luciferase (Gluc).
- Author:
Ling ZHANG
;
Xiaoming LIU
;
Jingdong SONG
;
Yan XIN
;
Yao DENG
;
Wenjie TAN
- Publication Type:Journal Article
- MeSH:
Animals;
Copepoda;
Genes, Reporter;
Genetic Vectors;
genetics;
metabolism;
Hepacivirus;
genetics;
metabolism;
Hepatitis C;
virology;
Humans;
Lentivirus;
genetics;
metabolism;
Luciferases;
genetics;
metabolism;
Recombinant Fusion Proteins;
genetics;
metabolism;
Viral Structural Proteins;
genetics;
metabolism
- From:
Chinese Journal of Virology
2015;31(2):174-179
- CountryChina
- Language:Chinese
-
Abstract:
To develop a recombinant lentivirus co-expressing structural protein of hepatitis C virus (HCV) and secreted Gaussia Luciferase (Gluc), we first constructed an expression vector that encoded HCV structural protein (C, E1, E2) and GLuc named pCSGluc2aCE1E2. The expression of HCV proteins and Gluc was confirmed by an immunofluorescence assay (IFA) and the detection of luciferase activity. Recombinant lentivirus (VSVpp-HCV) was developed by the co-transfection of pCSGluc2aCE1E2 into 293T cells with pHR'CMVA8.2 and pVSVG. The infectivity of VSVpp-HCV was confirmed by luciferase activity detection, IFA and western blotting. Virus-like particles were identified using electron microscopy after concentration. The results showed that the level of luciferase activity correlated with the expression of HCV protein after the infection of cells with lentivirus VSVpp-HCV. Therefore, the expression level of HCV proteins could be evaluated by detecting the luciferase activity of Gluc. In conclusion, this research pave a way for the development of transgenic mice that express HCV proteins and Gluc, which enable the evaluation of anti-HCV therapy and vaccine in vivo.
