SENSING SUPERFUND CHEMICALS WITH RECOMBINANT SYSTEMS

某些细菌含有编码蛋白质和/或酶的基因， 其降解或赋予对有毒化合物的抗性。 将从这些质粒构建生物发光报告质粒。 基因（含其启动子元件）与其他基因连锁 具有发光能力（细菌荧光素酶基因）。 这些质粒将被引入合适的宿主细菌中， 以产生能够检测低浓度的生物检测系统， 铬等超级基金有害物质的浓度 (VI)砷（砷酸盐和亚砷酸盐）和镍。 重组 将检查细菌的抗性基因表达， 荧光素酶基因在有害物质的存在。 具体地，当目标有害物质存在于环境中时， 样品，从基因中产生抗性相关蛋白， 诱导构建的报告质粒。 因为相同的 质粒含有荧光素酶基因，荧光素酶也将被 细菌就会发光。这种生物发光是 与目标危险化学品的存在直接相关 并构成分析信号。 基因设计的 细菌将被用于生物传感器的开发， 检测系统。 通过将这些细菌放置在 纤维高灵敏度和选择性的生物传感器将被开发 能够监测超级基金有害物质的水平， 环境样品（地下水和土壤）。 建设 传感器将在光学设计方面被优化， 细菌选择和细菌固定化。 等因素 响应时间、选择性和固体表面的影响 将对土壤进行评估。 进一步的评估将由 在实际超级基金样本中测试传感器响应， 使用传统的提取方法对污染物进行定量， 光谱技术

Certain bacteria contain genes that encode proteins and/or enzymes, which degrade or confer resistance to toxic compounds. Bioluminescent reporter plasmids will be constructed from these genes (containing their promoter elements) by linkage to other genes with the capability of light emission (bacterial luciferase genes). These plasmids will be introduces into suitable host bacteria in order to produce a biological detection system, capable of detecting low concentrations of Superfund hazardous substances such as chromium (VI), arsenic (arsenate and arsenite), and nickel. The recombinant bacteria will be examined for expression of the resistance genes and the luciferase genes in the presence of the hazardous substances. Specifically, when the targeted hazardous substance is present in the sample, production of resistance-related proteins from the genes in the constructed reported plasmid will be induces. Since the same plasmid contains the luciferase genes, luciferase will also be produced and the bacteria will bioluminesce. This bioluminescence is directly related to the presence of the targeted hazardous chemical and constitutes the analytical signal. The genetically designed bacteria will be employed in the development of biosensor based detection systems. By positioning these bacteria in front of optical fibers highly sensitive and selective biosensors will be developed capable of monitoring the level of Superfund hazardous substances in environmental samples (groundwater and soil). The construction of the sensors will be optimized with respect to the optical design, bacteria selection, and bacteria immobilization. Factors such as response time, selectivity, and effect of the solid surface in the case of soil will be evaluated. Further evaluation will be carried out by testing the sensor response in actual Superfund samples and quantification of the contaminants using traditional extraction and spectroscopic techniques.