绿色荧光蛋白(GFP)基因在短小芽孢杆菌中的组成型表达  

Expression of a Green Fluorescent Protein in Rice Epiphyte Bacillus brevis Under the Direction of a Constitutive Promoter Functional Fragment F1

作　　者：陈云鹏[1] 曲志才[1] 严健[1] 王敬文[1] 叶鸣明[1] 沈大棱[1] 

机构地区：[1]复旦大学生命科学学院遗传学研究所基因多样性与设计农业研究中心,上海200433

出　　处：《复旦学报（自然科学版）》2003年第4期541-545,共5页Journal of Fudan University：Natural Science

基　　金：美国McKnight基金资助项目(14001404);上海市科委资助项目(KBH1322093);上海高校优秀学生"创业浦东直通车"课题

摘　　要：采用PCR技术,亚克隆来自短小芽孢杆菌总基因组中的启动子活性片段F1,构建了一个大肠杆菌 短小芽孢杆菌穿梭表达载体pHY300 F1gfp,以缺失启动子的绿色荧光蛋白(GFP)基因为报告基因,检测了该片段在短小芽孢杆菌DX01菌株中的启动活性,在荧光显微镜下,观察到了明亮的绿色荧光,证实活性片段F1具有组成型启动子的功能,GFP基因在短小芽孢杆菌中实现了组成型表达.The green fluorescent protein(GFP) of the jellyfish Aequorea Victoria is a useful reporter molecule for monitoring gene expression in vivo in eukaryotic and prokaryotic cells. A GFP vector was constructed for in situ detection of the rice epiphyte Bacillus brevis strain DX01. The gfpS65T gene was transcriptionally fused to a strong B. brevis promoter functional fragment F1,which was subcloned from the genome of strain DX01 by PCR technique, and then inserted into an Escherichia coliBacillus shuttle vector pHY300PLK. DX01 cells harboring pHY300F1gfp could produce bright green fluorescence when they were examined by using fluorescent microscopy. The results were confirmed by western blotting and fluorescenceactivated cell sorting (FACS). The promoter Functional fragment F1 showed constitutive activity.

关 键 词：绿色荧光蛋白 短小芽孢杆菌 启动子活性片段 组成型表达 

分 类 号：Q786[生物学—分子生物学]