Department of Civil and Environmental Engineering, Seoul National University, Gwanakno 599, Seoul 151-742, Republic of Korea.
Environ Pollut. 2011 Feb;159(2):509-14. doi: 10.1016/j.envpol.2010.10.017. Epub 2010 Nov 18.
A phenanthrene-degrading bacterium, Sphingomonas paucimobilis EPA505 was used to construct two fluorescence-based reporter strains. Strain D harboring gfp gene was constructed to generate green fluorescence when the strain started to biodegrade phenanthrene. Strain S possessing gef gene was designed to die once phenanthrene biodegradation was initiated and thus to lose green fluorescence when visualized by a live/dead cell staining. Confocal laser scanning microscopic observation followed by image analysis demonstrates that the fluorescence intensity generated by strain D increased and the intensity by strain S decreased linearly at the phenanthrene concentration of up to 200 mg/L. Such quantitative increase and decrease of fluorescence intensity in strain D (i.e., from 1 to 11.90 ± 0.72) and strain S (from 1 to 0.40 ± 0.07) were also evident in the presence of Ottawa sand spiked with the phenanthrene up to 1000 mg/kg. The potential use of the reporter strains in quantitatively determining biodegradable or toxic phenanthrene was discussed.
一株菲降解菌，即少动鞘氨醇单胞菌 EPA505，被用于构建两种基于荧光的报告菌株。构建携带 GFP 基因的菌株 D 是为了当该菌株开始降解菲时产生绿色荧光。设计携带 gef 基因的菌株 S 在菲生物降解开始时死亡，因此在用活/死细胞染色法观察时失去绿色荧光。共聚焦激光扫描显微镜观察和图像分析表明，在菲浓度高达 200mg/L 的情况下，菌株 D 产生的荧光强度增加，而菌株 S 的荧光强度线性降低。在含有菲的渥太华砂中，菌株 D（从 1 增加到 11.90±0.72）和菌株 S（从 1 减少到 0.40±0.07）的荧光强度也出现了这种定量增加和减少。报告菌株在定量测定可生物降解或有毒菲方面的潜在用途进行了讨论。
