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Final Progress Reports: University of Kentucky: Sensing Superfund Chemicals with Recombinant Systems

Superfund Research Program

Sensing Superfund Chemicals with Recombinant Systems

Project Leaders: Sylvia Daunert (University of Miami), Sylvia Daunert (University of Miami)
Co-Investigators: Leonidas G. Bachas (University of Miami), Leonidas G. Bachas (University of Miami)
Grant Number: P42ES007380
Funding Period: 1997-2014

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Final Progress Reports

Year:   2013  2007  2004  1999 

The objective of the team’s research is to develop sensing systems employing genetically engineered bacteria that produce a measurable signal in the presence of Superfund chemicals.  Furthermore, the researchers have demonstrated the application of these bacterial sensing systems in monitoring real environmental samples and adaptation of this system to micro-total analytical systems (µTas).  Genetically engineered whole cell biosensors are designed by introducing in a living cell a reporter gene whose expression is activated as a consequence of the interaction between the target compound and a regulatory protein.  This interaction subsequently activates expression of the reporter gene through the promoter (O/P) sequence.  The signal provided by the reporter protein can be measured and directly related to the target compound concentration.  On the basis of this strategy the researchers have developed sensing systems for chlorocatechol, arsenic, biphenyl, and hydroxy-PCBs. 

Validation of the team’s assays for arsenic and chlorocatechols was accomplished by employing them in monitoring real environmental samples.  With the goal of developing systems that are amenable for field analysis and high throughput of samples, the team integrated a cell-based assay for arsenic detection using green fluorescent protein (GFP) as a reporter into a microcentrifuge microfluidics platform.  Using this platform, they were able to detect levels of arsenic within the standards set by EPA.  The assay was performed with minimal induction time (< 1min), allowing for faster analysis.  The whole cell-based assay demonstrated to be highly sensitive and selective, thus allowing for the measurement of lower levels of pollutants as compared to conventional methods of analysis.  Furthermore, the reagents employed in these assays are bacteria that can be prepared on a large scale at low cost.  Thus, the team’s whole cell assays are cost-effective, amenable to miniaturization, rugged, and suited for applications in a variety of field samples.

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