title: [Construction of an expression vector with a bicistron of green fluorescent protein and HCV 5'NCR controlled luciferase gene and its expression in hepatocytes].
abstract: 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.
authors.affiliation: Institute of Infectious Disease, Xiangya Hospital, Department of Microbiology, Xiangya School of Medicine, Central South University, Changsha 410008, China.
authors.forename: Shui-ping
authors.initials: SP
authors.lastname: Liu
authors.forename: De-ming
authors.initials: DM
authors.lastname: Tan
authors.forename: Zhou-hua
authors.initials: ZH
authors.lastname: Hou
mesh_terms: 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
chemical_list: 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