Cloning and mutagenetic modification of the firefly luciferase gene and its use for bioluminescence microscopy of expression during metamorphosis.

Bioluminescence microscopy is an area attracting considerable interest in the field of cell biology because it offers several advantages over fluorescence microscopy, including no requirement for excitation light and being phototoxicity free. This method requires brighter luciferase for imaging; however, suitable genetic resource material for this purpose is not available at present. To achieve brighter bioluminescence microscopy, we developed a new firefly luciferase. Using the brighter luciferase, a reporter strain of Gal4-UAS (Upstream Activating Sequence) system was constructed. This system demonstrated the expression pattern of , which is a segment polarity gene, during metamorphosis by bioluminescence microscopy, and revealed drastic spatiotemporal change in the expression pattern during head eversion in the early stage of pupation.