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Limitations of the reporter green fluorescent protein under simulated tumor conditionsCoralli, Claudia ...This paper reports a detailed analysis of the effect of low oxygen conditions (hypoxia) on the reporter green tiuorescent protein (GFP). It questions the feasibility of using GFP for gene expression ... studies under tumor conditions. Hypoxia is a characteristic of both experimental and clinical tumors. Several important factors are pointed out which need to be considered when using GFP as reporter gene. GFP tluorescence is the final product of a long and complex pathway involving transcription, translation, and posttranslational modifications. All of these steps may be affected by the availability of oxygen. We show specifically that cellular GFP fluorescence decreased with reduced oxygenation, anoxia virtually eliminated tluorescence and protein levels, and fluorescence recovery after anoxia required 5-10 h of reoxygenation. In conclusion, GFP appears to be a good marker gene to study location or movement of proteins or cells but should be used with great caution as a reporter of gene expression under tumor conditions.Source: Cancer research. - ISSN 0008-5472 (Letn. 61, št. 12, 2001, str. 4784-4790)Type of material - article, component partPublish date - 2001Language - englishCOBISS.SI-ID - 14546905
Author
Coralli, Claudia |
Čemažar, Maja |
Kanthou, Chryso |
Tozer, Gillian M. |
Dachs, Gabi U.
Topics
Bladder neoplasms |
Metabolism |
Genetics |
Genes, reporter |
Luminescent proteins |
Metabolism |
Biosynthesis |
Oxygen |
Metabolism |
Mice, scid |
Cell hypoxia |
Fluorescence |
Gene expression |
Indicators and reagents |
Rna, messenger |
Biosynthesis |
Metabolism |
Transfection |
Tumor cells, cultured |
Geni, poročevalci |
Kisik |
Luminiscentne beljakovine |
Sečni mehur, novotvorbe |
Celična hipoksija |
Fluorescenca |
Genetsko izražanje |
Indikatorji in reagenti |
Miši, mutiranih sevov |
RNA, prenašalna |
Transfekcija |
Tumorske celice, kulture
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