【摘要】 以Escherichiacoli高效表达载体pET-29a为基础,将广宿主载体pUCP19中稳定的假单胞菌(Pseudomonas)质粒DNA复制子和自行分离克隆的荧光假单胞菌(P.fluorescens)P303组成型表达启动子PP303 插入其中,分别获得了重组载体pQMV(4.6 kb)和pGMP(5.6 kb)。它们均含有假单胞菌和大肠杆菌复制子、荧光假单胞菌内源强启动子,以及包含8~11个限制性内切酶多克隆酶切位点;此外,载体pGMP还含有绿色荧光蛋白基因gfp作为辅助检测标记。连续培养和继代培养测定,这两种载体在荧光假单胞菌中的稳定性均为100%(120 h)。更多还原

【Abstract】 The replicon of Pseudomonas plasmid DNA from pUCP19, and a new strong promoter PP303, cloned from Pseudomonas fluorescens P303 strain, were inserted into plasmid pET-29a, a high level expression vector of Escherichia coli. The recombinant shuttle expression plasmid pQMV(4.6 kb) and pGMP(5.6 kb) were obtained respectively. The vector pGMP containing gfp gene was especially available for detection and monitoring. The stability of the two expression vectors were 100%(120 h) in P303 strain of Pseudomonas fluorescens. Both recombinant vectors will be powerful for research on gene expression, regulation and construction of engineered strains of Pseudomonas.更多还原