Development of a p53 responsive GFP reporter; identification of live cells with p53 activity.
p53 is among the most intensely studied human proteins because of its vital role as the prototype tumor suppressor. As a result, there are widespread applications for p53 functional analysis in biotechnology as it relates to cancer research. p53 is a potent sequence specific transcription factor, which induces the expression of a number of genes whose products mediate cell cycle arrest and apoptosis. Because the tumor suppressor activity of p53 is dependent on its transcription regulatory function, we have undertaken to develop a p53-responsive green fluorescent protein reporter strategy to enable the identification of live cells containing p53 transcriptional transactivation activity. We demonstrate within the use of GFP fluorescence to monitor both endogenous and plasmid derived p53 biochemical and biological activity. Identifying live cells with p53 activity through GFP fluorescence will have wide application for both in vitro and in vivo studies of the p53 tumor suppressor protein.
Zhang
W W
WW
Department of Microbiology and Immunology, McGill University, 3775 University Street, Que., H3A 2B4, Montreal, Canada.
Labrecque
S
S
Azoulay
E
E
Dudley
R
R
Matlashewski
G
G
D017209:Apoptosis / Q000502:physiology; D015972:Gene Expression Regulation, Neoplastic; D017930:Genes, Reporter / Q000235:genetics*; D049452:Green Fluorescent Proteins; D006801:Humans; D007202:Indicators and Reagents / Q000378:metabolism*; D008164:Luminescent Proteins / Q000235:genetics*; D012516:Osteosarcoma; D014158:Transcription, Genetic / Q000502:physiology; D014162:Transfection; D014407:Tumor Cells, Cultured / Q000166:cytology / Q000502:physiology; D016159:Tumor Suppressor Protein p53 / Q000235:genetics*
D007202:Indicators and Reagents; D008164:Luminescent Proteins; D016159:Tumor Suppressor Protein p53; D049452:Green Fluorescent Proteins