Departments of Plant and Soil Science and Molecular and Cell Biology, University of Aberdeen, Aberdeen AB9 2UE, Scotland.
Appl Environ Microbiol. 1992 Aug;58(8):2444-8. doi: 10.1128/aem.58.8.2444-2448.1992.
Genes encoding bioluminescence from Vibrio harveyi were cloned into Pseudomonas syringae pv. phaseoli-cola, resulting in high levels of bioluminescence. After inoculation of sterile and nonsterile soil slurries with bioluminescent P. syringae, cells could not be identified by conventional light microscopy. However, when we used charge coupled device-enhanced microscopy, bioluminescent single cells were detected easily in dark fields despite masking by soil particulate matter, and in addition, the extent of competition from indigenous soil bacteria could be monitored. The technique which we describe offers great potential for tracking and determining the spatial distribution of genetically marked microorganisms in the environment.
从哈维氏弧菌中克隆出编码生物发光的基因，并将其插入到丁香假单胞菌 pv. phaseoli-cola 中，从而实现高水平的生物发光。将生物发光的荧光假单胞菌接种到无菌和非无菌土壤悬浮液中后，用传统的光学显微镜无法识别细胞。但是，当我们使用电荷耦合器件增强显微镜时，即使被土壤颗粒物质掩盖，也可以轻松地在暗场中检测到生物发光的单细胞，此外，还可以监测土著土壤细菌的竞争程度。我们所描述的这项技术具有很大的潜力，可以跟踪和确定环境中遗传标记微生物的空间分布。
