Ogoh Katsunori, Akiyoshi Ryutaro, Suzuki Hirobumi
Olympus Corporation, Kuboyama 2-3, Hachioji, Tokyo, 192-8512, Japan.
Biochem Biophys Rep. 2020 May 30;23:100771. doi: 10.1016/j.bbrep.2020.100771. eCollection 2020 Sep.
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.
生物发光显微镜是细胞生物学领域中备受关注的一个领域，因为它相对于荧光显微镜具有几个优势，包括不需要激发光且无光毒性。这种方法需要更亮的荧光素酶用于成像；然而，目前尚无适用于此目的的遗传资源材料。为了实现更亮的生物发光显微镜成像，我们开发了一种新的萤火虫荧光素酶。利用这种更亮的荧光素酶，构建了Gal4-UAS（上游激活序列）系统的报告菌株。该系统通过生物发光显微镜展示了一种节段极性基因在变态过程中的表达模式，并揭示了在化蛹早期头部外翻期间该基因表达模式的剧烈时空变化。
