Genome engineering technologies have rapidly advanced the speed at which cellular genotypes can be altered. However, from a synthetic biological perspective, we often lack the assays necessary to connect phenotype with genotype. This is a common challenge in metabolic engineering, which is often hampered by the low throughput of analytical chemistry techniques. Fluorescent biosensors can, in principle, provide a rapid, single-cell quantification of metabolic state but lack generality and have historically been difficult to construct. Here we describe a methodology – which we term domain insertion profiling - for the rapid construction and isolation of fluorescent biosensors using the green fluorescent protein and any ligand-binding protein domain. As a proof of concept, we demonstrate the isolation, optimization, and function of several biosensors against central carbon intermediates and discuss the general utility and application of biosensors in synthetic biology.

Rapid construction of metabolite biosensors using domain insertion profiling

251st National Meeting (2016)

The official archive and repository for all ACS Meetings & Expositions abstracts and select posters or presentations.