Vollmer A C, Belkin S, Smulski D R, Van Dyk T K, LaRossa R A
Swarthmore College, Pennsylvania 19081-1397, USA.
Appl Environ Microbiol. 1997 Jul;63(7):2566-71. doi: 10.1128/aem.63.7.2566-2571.1997.
Plasmids were constructed in which DNA damage-inducible promoters recA, uvrA, and alkA from Escherichia coli were fused to the Vibrio fischeri luxCDABE operon. Introduction of these plasmids into E. coli allowed the detection of a dose-dependent response to DNA-damaging agents, such as mitomycin and UV irradiation. Bioluminescence was measured in real time over extended periods. The fusion of the recA promoter to luxCDABE showed the most dramatic and sensitive responses. lexA dependence of the bioluminescent SOS response was demonstrated, confirming that this biosensor's reports were transmitted by the expected regulatory circuitry. Comparisons were made between luxCDABE and lacZ fusions to each promoter. It is suggested that the lux biosensors may have use in monitoring chemical, physical, and genotoxic agents as well as in further characterizing the mechanisms of DNA repair.
构建了质粒，其中来自大肠杆菌的DNA损伤诱导型启动子recA、uvrA和alkA与费氏弧菌luxCDABE操纵子融合。将这些质粒导入大肠杆菌后，可检测到对丝裂霉素和紫外线照射等DNA损伤剂的剂量依赖性反应。在较长时间内实时测量生物发光。recA启动子与luxCDABE的融合显示出最显著和敏感的反应。证明了生物发光SOS反应对lexA的依赖性，证实了该生物传感器的报告是通过预期的调控电路传递的。对每个启动子的luxCDABE和lacZ融合进行了比较。有人提出，lux生物传感器可用于监测化学、物理和遗传毒性剂，以及进一步表征DNA修复机制。
