Science. 1990 Aug 17;249(4970):778-81. doi: 10.1126/science.249.4970.778.
A bioluminescent reporter plasmid for naphthalene catabolism (pUTK21) was developed by transposon (Tn4431) insertion of the lux gene cassette from Vibrio fischeri into a naphthalene catabolic plasmid in Pseudomonas fluorescens. The insertion site of the lux transposon was the nahG gene encoding for salicylate hydroxylase. Luciferasemediated light production from P. fluorescens strains harboring this plasmid was induced on exposure to naphthalene or the regulatory inducer metabolite, salicylate. In continuous culture, light induction was rapid (15 minutes) and was highly responsive to dynamic changes in naphthalene exposure. Strains harboring pUTK21 were responsive to aromatic hydrocarbon contamination in Manufactured Gas Plant soils and produced sufficient light to serve as biosensors of naphthalene exposure and reporters of naphthalene biodegradative activity. The robust and sensitive nature of the bioluminescent reporter technology suggests that new sensing methods can be developed for on-line process monitoring and control in complex environmental matrices.
构建了一个用于萘降解的生物发光报告质粒（pUTK21），它是通过转座子（Tn4431）将发光菌（Vibrio fischeri）的 lux 基因盒插入到荧光假单胞菌（Pseudomonas fluorescens）的萘降解质粒中而得到的。lux 转座子的插入位点是编码水杨酸羟化酶的 nahG 基因。当含有该质粒的荧光假单胞菌菌株暴露于萘或调节诱导代谢物水杨酸时，会诱导产生生物发光。在连续培养中，光诱导非常迅速（15 分钟），并且对萘暴露的动态变化高度敏感。携带 pUTK21 的菌株对煤气厂土壤中的芳烃污染有反应，并产生足够的光作为萘暴露的生物传感器和萘生物降解活性的报告器。生物发光报告基因技术的稳健性和敏感性表明，可以为复杂环境基质中的在线过程监测和控制开发新的传感方法。
