( ther development of the luciferase assa...)Genetic Reporter Systems

ther development of the luciferase assay, cells are co-transfected with a control plasmid of a different luciferase from renilla (Renilla reniformis). The activities of firefly and renilla luciferase can be measured separately in one sample. The activity of the renilla luciferase can therefore be used as an internal control for comparing different transfection experiments. Both luciferases are also used in co-transfection experiments for the parallel examination of two cis elements.

Green Fluorescent Protein (GFP)

Unlike other bioluminescent reporters, the green fluorescent protein from the Aequorea victoria jellyfish requires no additional proteins, substrates or co-factors to emit light. When irradiated with UV light or blue light, it emits green light, which enables the examination of gene expression and protein localization in situ and in vivo. In addition, the gene expression can be observed in real time. However, the system is less suitable for quantifying the gene expression.

Variations of GFP with different absorption and emission maxima also enable double-labeling experiments, such as the simultaneous examination of gene expression of two promoters in one cell, or the location of two different fusion proteins in one cell. Other variations of GFP are particularly designed for expression in mammalian cells or have up to a 35-fold higher fluorescence. With the aid of a GFP system with a drastically reduced half-life, dynamic processes can also be examined in vivo in the cell.

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