LOW-LIGHT IMAGE-ANALYSIS OF TRANSGENIC ORGANISMS USING BACTERIAL LUCIFERASE AS A MARKER

Methods for measurement of a novel light-emitting reporter gene system in bacteria, yeast, plant cells, plant tissues and intact plant organ s are described. The principle underlying the assay procedures is the bacterial luciferase catalysed oxidation of reduced flavin mononucleot ide (FMNH(2)) in the presence of the ten carbon aldehyde decanal, to y ield FMN, decanoic acid, water and a photon of light at 490 nm which c an be captured by X-ray film, a photomultiplier tube or, for in vivo m easurements, an image-intensifier coupled to a video camera. This ligh t measuring assay system is sensitive, easy to use, inexpensive, does not require radioactivity, and has been used successfully for rapid de tection of bacterial transformants, the quantitative measurement of tr ansient and stable gene expression in bacteria and yeast, and in vivo measurement of temporal and spatial gene expression throughout plant a nd animal development.