Bioluminescent Detection of Chemical Agent Contamination

Chemical weapons such as organophosphate (OP) nerve agents pose a deadly risk to both military and civilian personnel as exemplified by the use of nerve agents during the Iran/Iraq war and the Tokyo terrorist subway attack of 1995 which caused over 3,000 casualties. In addition, OP pesticide contamination of soils and water supplies is thought to result in 1 to 3 million cases of pesticide poisonings annually worldwide. Consequently, there is an urgent need for natural biological systems that can detect and safely dispose of these toxic compounds. This proposal will generate a genetically engineered yeast biosensor that can detect and hydrolyze chemical warfare agents such as VX. Importantly, the yeast biosensor will also be able to detect the subsequent biodegraded products. The yeast Saccharomyces cerevisiae, which is a robust non-pathogenic microorganism that is resistant to environmental extremes, will be used as a self-contained biosensor. No exogenous substrates or consumables will be required and a simple hand held illumination device will allow distinct visual detection of the chemical agent contamination and biodegraded products using a dual fluorescent reporter system.