Essential (and Dispensable) Genes in Escherichia Coli

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This project utilizes Tn5 derived DNA transposition systems to address several fundamental questions in bacterial genomics. The experiments will define the compliment of essential, dispensable and conditionally essential genes in the Escherichia coli K12 genome. This effort will provide a powerful tool for studying the function of genes of unknown function. In addition the E. coli K12 gene content will be reduced to the minimal necessary for survival for aerobic growth in rich medium. This will result in the generation of an organism that is more amenable to metabolic modeling. Finally, the techniques will be used as a tool for studying in vivo chromosome condensation. The project has broad impacts beyond basic science discoveries. The methodologies to be employed can be used to study any bacterial species amenable to DNA electroporation technology and thus can be used, for instance to identify functions necessary for pathogen survival in host organisms. In addition, previous work on this technology has resulted in the generation of a number of patents that have been licensed to a biotechnology supply company. These types of practical discoveries will continue. Finally, the research program has always been used as a vehicle for undergraduate education especially in regards to underrepresented minorities. This has been accomplished by having undergraduates as active participants in the research projects.