Abstract
Bioassays for toxic effects in mammals require the use of appropriate reporters in mammalian cell systems. Fluorescent Proteins of different colors offer the possibility to study different processes in the same living cell. Here the suitability of a separate and combined measurement of Enhanced Green Fluorescent Protein (EGFP) and the recently available Red Fluorescent Protein (DsRed) from Discosoma sp. was examined in order to create a combined bioassay for gene expression and cytotoxic effects. In stably transfected EGFP-expressing Chinese Hamster Ovary cells, the whole population shows strong green fluorescence, while stable transfection with pDsRed-N1 resulted in DsRed-expressing colonies in which red fluorescence was restraint to their center. DsRed-expressing cell lines with 90 % DsRed expression were isolated. EGFP and DsRed can be differentiated and quantified by microplate reader measurements (filter combinations 460/40+508/20 and 540/25+590/35) and FACS analysis (argon laser 488 nm). Thus, they are suitable for construction of a two-parameter-bioassay.
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Baumstark-Khan, C., Hellweg, C.E., Horneck, G. (2002). On the Suitability of Red and Enhanced Green Fluorescent Protein (DsRed/EGFP) as Reporter Combination. In: Hoffmann, KH. (eds) Coupling of Biological and Electronic Systems. Springer, Berlin, Heidelberg. https://doi.org/10.1007/978-3-642-56177-1_1
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DOI: https://doi.org/10.1007/978-3-642-56177-1_1
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