Bioluminescent reporter genes for promoter discovery

Abstract

Discovery of promoter elements with previously unknown regulated responses is important for metabolic engineering. For example, promoters responsive to the end product can be useful to regulate expression with increasing levels of product. In addition, such promoters can be used as screens for production strain with increased titers. Use of reporter genes, such as a bioluminescent reporter luxCDABE, can facilitate promoter discovery. Here, protocols for analysis of genome-wide luxCDABE reporter gene collections in Escherichia coli are provided. Further, a protocol for using a selected para-hydroxycinnamic (pHCA)-responsive promoter as detection assay for bioproduced pHCA is provided. © 2012 Springer Science+Business Media, LLC.