title: Labeling of human insulin-like growth factor-I eukaryotic expression vector with green fluorescent protein.
abstract: OBJECTIVE
  To label human insulin-like growth factor-I (hIGF-I) eukaryotic expression vector with green fluorescent protein (GFP) for the repair of articular cartilage defects.
  
  
  METHODS
  GFP cDNA was inserted into pcDNA(3.1)-hIGF-1 to construct the co-expression vector with two multiple cloning sites mammalian expression vector under two cytomegalovirus promoters/enhancers respectively. Recombinant pcGI was transfected into NIH 3T3 cells with the help of lipofectamine.
  
  
  RESULTS
  Enzyme digestion and agarose gel electrophoresis analysis revealed that pcGI vector contained correct GFP and hIGF-I cDNA. Expression of hIGF-1 and GFP was confirmed in transfected NIH 3T3 cells by immunocytochemical analysis and fluorescence microscopy.
  
  
  CONCLUSIONS
  hIGF-I eukaryotic expression vector has been successfully labeled with GFP.
authors.affiliation: Department of Orthopaedic Surgery, First Hospital, Jilin University, Changchun 130021, China.
authors.forename: Yi
authors.initials: Y
authors.lastname: Liu
authors.forename: Shao-kun
authors.initials: SK
authors.lastname: Zhang
authors.forename: Hong
authors.initials: H
authors.lastname: Wu
authors.forename: Yu-xing
authors.initials: YX
authors.lastname: Shan
authors.forename: Gang
authors.initials: G
authors.lastname: Wang
authors.forename: Xin-xiang
authors.initials: XX
authors.lastname: Xu
mesh_terms: D002358:Cartilage, Articular / Q000293:injuries; D002478:Cells, Cultured; D005057:Eukaryotic Cells / Q000166:cytology / Q000502:physiology; D005347:Fibroblasts / Q000166:cytology / Q000502:physiology; D005786:Gene Expression Regulation; D015316:Genetic Therapy / Q000379:methods*; D005822:Genetic Vectors; D049452:Green Fluorescent Proteins / Q000494:pharmacology*; D006801:Humans; D007150:Immunohistochemistry; D007334:Insulin-Like Growth Factor I / Q000235:genetics / Q000627:therapeutic use*; D049408:Luminescent Agents / Q000494:pharmacology; D012680:Sensitivity and Specificity; D014162:Transfection / Q000379:methods*; D014947:Wounds and Injuries / Q000175:diagnosis / Q000628:therapy
chemical_list: D049408:Luminescent Agents; D049452:Green Fluorescent Proteins; D007334:Insulin-Like Growth Factor I