Dollard Marie Andrée, Billard Patrick
U.R. Ecotoxicité, Biodiversité, Santé Environnementale, Université de Metz, Campus Bridoux-Rue du Général Delestraint, 57070, Metz, France.
J Microbiol Methods. 2003 Oct;55(1):221-9. doi: 10.1016/s0167-7012(03)00164-7.
A phosphate sensor plasmid was constructed, in which the inducible promoter of the alkaline phosphatase gene (phoA) from Escherichia coli is fused to the bioluminescence genes from Vibrio fischeri. The reporter construct was introduced into E. coli MG1655 and the rhizosphere coloniser Pseudomonas fluorescens DF57, which produced light in a dose-dependent manner when exogenous phosphate concentrations fell below 60 and 40 microM, respectively. These strains also responded to various organic and inorganic phosphorus compounds. Their ability to distinguish the bioavailable portion of phosphate in standard solution was demonstrated using different phosphate ligands. When applying the bioassay to wastewater samples, luminescence patterns correlated with phosphate concentrations determined by standard chemical procedure. These results indicated that phoA::lux-based bacterial sensors may serve as tools for the assessment of phosphate bioavailability.
构建了一种磷酸盐传感器质粒,其中来自大肠杆菌的碱性磷酸酶基因(phoA)的诱导型启动子与费氏弧菌的生物发光基因融合。将报告构建体导入大肠杆菌MG1655和根际定殖菌荧光假单胞菌DF57,当外源磷酸盐浓度分别降至60和40微摩尔以下时,它们会以剂量依赖的方式发光。这些菌株也对各种有机和无机磷化合物有反应。使用不同的磷酸盐配体证明了它们区分标准溶液中磷酸盐生物可利用部分的能力。当将生物测定应用于废水样品时,发光模式与通过标准化学程序测定的磷酸盐浓度相关。这些结果表明,基于phoA::lux的细菌传感器可作为评估磷酸盐生物可利用性的工具。