Labeling of human insulin-like growth factor-I eukaryotic expression vector with green fluorescent protein.
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.
Liu
Yi
Y
Department of Orthopaedic Surgery, First Hospital, Jilin University, Changchun 130021, China.
Zhang
Shao-kun
SK
Wu
Hong
H
Shan
Yu-xing
YX
Wang
Gang
G
Xu
Xin-xiang
XX
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
D049408:Luminescent Agents; D049452:Green Fluorescent Proteins; D007334:Insulin-Like Growth Factor I