Sunday, May 4, 2008
2-55

A plasmid-based system for the use in coupling non-selectable phenotypes to selectable phenotypes

Paul Handke, Chemical and Biological Engineering, University of Colorado, 1111 Engineering Drive, UCB 424, Boulder, CO 80309, Michael D. Lynch, OPX Biotechnologies, Inc., 5541 Central Ave., Suite 270, Boulder, CO 80301, and Ryan T. Gill, Department of Chemical and Biological Engineering, University of Colorado, 1111 Engineering Drive, UCB 424, Boulder, CO 80309.

The production of high value commodity chemicals and bio-fuels by microorganisms relies on the identification of genes in mutants that allow for increased production. Having a reporter system that directly responds to the desired product would enable easier screens and/or selections of said mutants. However, most naturally occurring reporters, such as the Escherichia coli lactose promoter, are often sub-optimal as they may express at low levels, exhibit loose regulation, and respond only in natural conditions or environments. The goal of this work is therefore to develop a plasmid-based approach to couple the non-selectable phenotype of chemical production to a selectable phenotype. Using both computational and experimental techniques, we have developed a reporter system in which the selectable response conferred is dependent upon inducer concentration. Development of such plasmids for the improved production of organic acids is the focus of current activities.