Kim June-Hyung, Roh Changhyun, Lee Chang-Won, Kyung Dohyun, Choi Soo-Keun, Jung Heung-Chae, Pan Jae-Gu, Kim Byung-Gee
Department of Chemical Engineering, Dong-A University, Busan 604-714, Korea.
J Microbiol Biotechnol. 2007 Apr;17(4):677-80.
To analyze a cotG-based Bacillus subtilis spore display system directly, GFP(uv) was expressed on the surface of Bacillus subtilis spores. When GFP(uv) was fused to the C-terminal of the cotG structural gene and expressed, the existence of a CotG-GFP(uv) fusion protein on the B. subtilis spore was confirmed by flow cytometry confocal microscopic analysis. When the cotG anchoring motif was deleted, no fluorescence emission was observed under flow cytometry and confocal microscopic analysis from the purified spore, confirming the essential role of CotG as an anchoring motif. This GFP(uv) displaying spore might be used for another signaling application triggered by intracellular or extracellular stimuli.
为了直接分析基于cotG的枯草芽孢杆菌孢子展示系统,绿色荧光蛋白(uv)(GFP(uv))在枯草芽孢杆菌孢子表面表达。当GFP(uv)与cotG结构基因的C末端融合并表达时,通过流式细胞术共聚焦显微镜分析证实了枯草芽孢杆菌孢子上存在CotG-GFP(uv)融合蛋白。当cotG锚定基序被删除时,在流式细胞术和共聚焦显微镜分析下,从纯化的孢子中未观察到荧光发射,这证实了CotG作为锚定基序的重要作用。这种展示GFP(uv)的孢子可用于由细胞内或细胞外刺激触发的另一种信号应用。