Labeling of human insulin-like growth factor-I eukaryotic expression vector with green fluorescent protein.
- Author:
Yi LIU
1
;
Shao-kun ZHANG
;
Hong WU
;
Yu-xing SHAN
;
Gang WANG
;
Xin-xiang XU
Author Information
- Publication Type:Journal Article
- MeSH: Cartilage, Articular; injuries; Cells, Cultured; Eukaryotic Cells; cytology; physiology; Fibroblasts; cytology; physiology; Gene Expression Regulation; Genetic Therapy; methods; Genetic Vectors; Green Fluorescent Proteins; pharmacology; Humans; Immunohistochemistry; Insulin-Like Growth Factor I; genetics; therapeutic use; Luminescent Agents; pharmacology; Sensitivity and Specificity; Transfection; methods; Wounds and Injuries; diagnosis; therapy
- From: Chinese Journal of Traumatology 2005;8(3):132-137
- CountryChina
- Language:English
-
Abstract:
OBJECTIVETo label human insulin-like growth factor-I (hIGF-I) eukaryotic expression vector with green fluorescent protein (GFP) for the repair of articular cartilage defects.
METHODSGFP 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.
RESULTSEnzyme 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.
CONCLUSIONShIGF-I eukaryotic expression vector has been successfully labeled with GFP.
