[Construction of an expression vector with a bicistron of green fluorescent protein and HCV 5'NCR controlled luciferase gene and its expression in hepatocytes].
OBJECTIVE
To construct an expression vector with a bicistron of green fluorescent protein (GFP) and HCV 5'NCR controlled luciferase gene and to explore its expression in hepatocytes.


METHODS
With molecular cloning techniques, the DNA fragment of HCV 5'NCR and its controlled luciferase gene was obtained from plasmid pCMVNCRLuc and subcloned into pEGFP-C1, an expression plasmid of GFP. A recombinant plasmid pGFPNCRLuc with a bicistron of GFP and HCV 5'NCR controlled luciferase gene was constructed, and liposome-mediated transient expression was detected in hepatocyte QSG7701 cells.


RESULTS
The recombinant plasmid expressed both GFP and luciferase at a high level. The expression activity of GFP showed no significant difference compared to that of pEGFP-C1 (P > 0.05), nor did that of luciferase to pCMVNCRLuc (P > 0.05).


CONCLUSIONS
An expression vector with a bicistron of GFP and HCV 5'NCR controlled luciferase gene was successfully constructed. GFP and luciferase could express independently. This study can promote the establishment of an anti-HCV drug screen system.
Liu
Shui-ping
SP
Institute of Infectious Disease, Xiangya Hospital, Department of Microbiology, Xiangya School of Medicine, Central South University, Changsha 410008, China.
Tan
De-ming
DM
Hou
Zhou-hua
ZH
D017930:Genes, Reporter / Q000235:genetics; D005822:Genetic Vectors; D049452:Green Fluorescent Proteins; D016174:Hepacivirus / Q000235:genetics*; D022781:Hepatocytes / Q000378:metabolism* / Q000821:virology; D006801:Humans; D008156:Luciferases / Q000096:biosynthesis / Q000235:genetics*; D008164:Luminescent Proteins; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000235:genetics; D014162:Transfection; D017361:Viral Nonstructural Proteins / Q000096:biosynthesis / Q000235:genetics*; D012324:RNA-Dependent RNA Polymerase
D049452:Green Fluorescent Proteins; D008156:Luciferases; D008164:Luminescent Proteins; D011994:Recombinant Proteins; D017361:Viral Nonstructural Proteins; C087036:NS-5 protein, hepatitis C virus; D012324:RNA-Dependent RNA Polymerase