Luciferase

Luciferase is an enzyme that emits light when supplied with one of its substrates, e.g. luciferin. This bioluminescence is used for several molecular approaches and the advantages are the fast and easy in vitro and in vivo detection, the high sensitivity as well as its non hazardous and non radioactive properties.

The pEGFPLuc-plasmid (Clontech) expressing fluorescence protein and the luciferase gene from firefly Photinus pyralis was stably transfected into different cell types with several methods. The fluorescent reporter gene is determined by flow cytometry a measuring the luciferase activity with the luciferase assay and the GloMax luminometer from Promega.

Cells transfected with pEGFPLuc (Clontech) are used for non-invasive in vivo imaging to monitor engraftment or metastasis of different cell types. An appropriate device (VisLuxxII, Visitron Systems) is available. Further on, pEGFPLuc can be used as co-marker to investigate transfection efficiencies.

HEK293 cells expressing the different reporter genes; magnification: 200x

Bioluminescence image from NMRI-nude mouse

HEK293 cells transfected with the pEGFPLuc-vector were
applied subcutaneously. Image was taken 41 days after cell injection.

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