摘要
应用DNA重组技术构建了表达绿色荧光蛋白 (GFP)的重组减毒鼠伤寒沙门氏菌。用EcoRⅠ和NcoⅠ将pEGFP质粒中的绿色荧光蛋白基因EGFP切下 ,克隆进表达载体质粒pYA3334的EcoRⅠ和NcoⅠ位点之间 ,构建成重组表达质粒pYAGFP ,然后转化大肠杆菌X6 2 12、中间宿主X3730、终末宿主X45 5 0。X45 5 0于LB培养基上呈绿色 ,在荧光显微镜下观察 ,整个菌体发绿色荧光。构建的表达GFP的重组鼠伤寒沙门氏菌为减毒沙门氏菌活载体基因工程疫苗的研制提供一个良好模型。
To provide a useful mode for development of recombinant Salmonella typhimurium vaccines, a recombinant avirulent S.typhimurium expressing green fluorescent protein (GFP) gene was constructed using recombinant DNA techniques. The GFP gene was excised from plasmid pEGFP and subcloned into expression vector pYA3334 as an EcoRⅠ/NcoⅠ fragment. The resulting recombinant plasmid was named as pYAGFP and then transformed into an avirulent S.typhimurium vaccine strain. The recombinant bacterial grew on LB plates as green colonies, which revealed green fluorescence under the fluorescent microscopy. These results suggest that The GFP gene was expressed correctly in the host bacterial and could be used for further studies on development of recombinant S.typhimurium vaccines.
基金
教育部高等学校骨干教师资助项目
