绿色荧光蛋白基因在木葡糖酸醋杆菌中的表达

摘要

Green fluorescent protein(GFP)as a reporter protein is widely used in signal conduction and gene expression and regulation in microbiology. In this research,GFPgenes were successfully expressed inGluconacetobacter xylinus with shut-tle plasmid pMV24. GFP genes were cloned from pMUTIN-gfp+ plasmid and connected to pMV24 to build prokaryotic ex-pression plasmid pMV24-gfp+. Electrotransformation was applied to introduce pMV24-gfp+ intoG. xylinus. The transformant exhibits bright green fluorescence when exposed to blue light in the ultraviolet range of fluorescence microscope. It can pro-vide very important theoretical basis for the observation of cell movement and the analysis of kinesin ofG. xylinus under chemotaxis.%绿色荧光蛋白(green fluorescent protein,GFP)广泛应用于报告基因、基因的表达与调控以及微生物信号传导.以pMV24穿梭质粒为载体,实现了GFP在木葡糖酸醋杆菌(Gluconacetobacter xylinus)中的成功表达.采用PCR技术扩增出绿色荧光蛋白基因,连接至木葡糖酸醋杆菌表达载体 pMV24 中,构建成功的质粒命名为 pMV24-gfp+.采用电转化技术将重组载体导入木葡糖酸醋杆菌中,转化子在荧光显微镜的蓝色激发光下发出绿色荧光,为木葡糖酸醋杆菌趋化性的观察及菌体中运动蛋白的分析提供了重要的理论依据.