Applegate B M, Kehrmeyer S R, Sayler G S
Department of Microbiology, University of Tennessee, Knoxville 37996-1605, USA.
Appl Environ Microbiol. 1998 Jul;64(7):2730-5. doi: 10.1128/AEM.64.7.2730-2735.1998.
A tod-luxCDABE fusion was constructed and introduced into the chromosome of Pseudomonas putida F1, yielding the strain TVA8. This strain was used to examine the induction of the tod operon when exposed to benzene, toluene, ethylbenzene, and xylene (BTEX) compounds and aqueous solutions of JP-4 jet fuel constituents. Since this system contained the complete lux cassette (luxCDABE), bacterial bioluminescence in response to putative chemical inducers of the tod operon was measured on-line in whole cells without added aldehyde substrate. There was an increasing response to toluene concentrations from 30 micrograms/liter to 50 mg/liter, which began to saturate at higher concentrations. The detection limit was 30 micrograms/liter. There was a significant light response to benzene, m- and p-xylenes, phenol, and water-soluble JP-4 jet fuel components, but there was no bioluminescence response upon exposure to o-xylene. The transposon insertion was stable and had no negative effect on cell growth.
构建了一个tod-luxCDABE融合基因，并将其导入恶臭假单胞菌F1的染色体中，得到菌株TVA8。该菌株用于检测暴露于苯、甲苯、乙苯和二甲苯（BTEX）化合物以及JP - 4喷气燃料成分的水溶液时tod操纵子的诱导情况。由于该系统包含完整的lux盒（luxCDABE），因此无需添加醛底物即可在全细胞中在线测量细菌对tod操纵子假定化学诱导剂的生物发光。对甲苯浓度从30微克/升增加到50毫克/升有递增响应，在更高浓度时开始饱和。检测限为30微克/升。对苯、间二甲苯和对二甲苯、苯酚以及水溶性JP - 4喷气燃料成分有显著的光响应，但暴露于邻二甲苯时没有生物发光响应。转座子插入是稳定的，对细胞生长没有负面影响。
