嗜麦芽假单胞菌热休克基因dnaK启动子的亚克隆及其在大肠杆菌中功能的研究

The subcloning of dnaK promoter from Pseudomonas maltophilia and its functions in E. coli

通过聚合酶链式反应 (PCR)得到了嗜麦芽假单胞菌 (Pseudomonasmaltophilia)热休克基因dnaK启动子的DNA片段dnaKp。将dnaKp的PCR的产物亚克隆在报道载体质粒 pUCD6 15的LuxCDABE基因上游 ,成重组的具有启动子的质粒融合体pUCD6 15 p。转化到大肠杆菌的pUCD6 15 p启动子对热、有机化合物和重金属的诱导响应快速灵敏 ,表现出强的启动子功能 ,能启动LuxCDABE基因表达 ,使荧光酶活性提高。借助荧光酶活性的变化 ,可检测系统中一些有机物和重金属的存在 。

The dnaK promoter from Pseudomonas maltophilia was obtained by PCR amplification and cloned at the point of upstream of LuxCDABE report gene vector, pUCD615. The plasmid borned dnaK::LuxCDABE fusant (pUCD615p) was transformed into E. coli , which showed strong responses to heat, organic compounds and heavy metals. As for the stress inducible expression system, luciferase activity went up at certain rate when the cells were exposed to stress. The recombinant E.coli strains allowed detection of stress producing environmental conditions by increasing light production.