Labeling of human insulin-like growth factor-I eukaryotic expression vector with green fluorescent protein

Chin J Traumatol. 2005 Jun;8(3):132-7.

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.

Publication types

  • Comparative Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Cartilage, Articular / injuries
  • Cells, Cultured
  • Eukaryotic Cells / cytology
  • Eukaryotic Cells / physiology
  • Fibroblasts / cytology
  • Fibroblasts / physiology
  • Gene Expression Regulation
  • Genetic Therapy / methods*
  • Genetic Vectors
  • Green Fluorescent Proteins / pharmacology*
  • Humans
  • Immunohistochemistry
  • Insulin-Like Growth Factor I / genetics
  • Insulin-Like Growth Factor I / therapeutic use*
  • Luminescent Agents / pharmacology
  • Sensitivity and Specificity
  • Transfection / methods*
  • Wounds and Injuries / diagnosis
  • Wounds and Injuries / therapy

Substances

  • Luminescent Agents
  • Green Fluorescent Proteins
  • Insulin-Like Growth Factor I