荧光蛋白在枯草芽孢杆菌中的表达  

Expression of fluorescent proteins in Bacillus subtilis

作　　者：刘敏胜[1] 韩冰[1] 邢新会[1] 

机构地区：[1]清华大学化学工程系,北京100084

出　　处：《清华大学学报（自然科学版）》2007年第6期874-877,共4页Journal of Tsinghua University(Science and Technology)

基　　金：国家自然科学基金资助项目(20176025;20336010);教育部博士点基金资助项目(20020003057)

摘　　要：枯草芽孢杆菌是生物工业用的重要宿主体系。为实现荧光蛋白从分子水平定量追踪枯草芽孢杆菌培养过程的目的,构建了一系列整合型表达载体pX-GFPmut1和游离型表达载体pSW1-GFPmut1,pSW1-CFP,pSW1-YFP,研究了不同颜色荧光蛋白在枯草芽孢杆菌中整合型表达和游离型表达的差异。最适合的荧光蛋白表达体系是用游离型质粒pSW1-GFPmut1在枯草芽孢杆菌中表达绿色荧光蛋白GFPmut1。含pSW1-GFPmut1枯草芽孢杆菌通过木糖诱导表达GFPmut1后,细胞破碎后的荧光强度和细胞浓度呈线性关系。结果表明,利用枯草芽孢杆菌表达荧光蛋白可以实现利用荧光蛋白快速定量枯草芽孢杆菌培养过程。Bacillus subtilis is an important host in bioindustry. A molecular-level quantification approach for bioprocesses using B. subtilis was developed using an integrated plasmid of pX-GFPmutl and non-integrated plasmids of pSW1-GFPmutl, pSW1-CFP, and pSW1-YFP to examine the expression of different color fluorescent proteins in Bacillus subtilis. The expression of GFPmutl via the nonintegrated plasmid of pSW1-GFPmutl was most suitable for Bacillus subtilis. The culture fluorescence after cell disruption by sonication showed a linear relationship with the cell concentration when the GFPmutl was expressed in B. subtilis by xylose induction. The results show that the expression of GFPmutl by pSW1-GFPmutl can be used for quantitative measurements for bioprocess detection and control of Bacillus subtilis.

关 键 词：荧光蛋白 快速定量 枯草芽孢杆菌 生物过程 

分 类 号：TQ920[轻工技术与工程—发酵工程]