A library of reporters of the global regulators of gene expression ofEscherichia coli

Author:

Dash SuchintakORCID,Jagadeesan RahulORCID,Baptista Ines S.C.ORCID,Chauhan VatsalaORCID,Kandavalli VinodhORCID,Oliveira Samuel M.D.ORCID,Ribeiro Andre S.ORCID

Abstract

SUMMARYThe topology of the transcription factor network (TFN) ofE. coliis far from uniform, with 22 global regulator (GR) proteins controlling one-third of all genes. So far, their production rates cannot be tracked by comparable fluorescent proteins. We developed a library of fluorescent reporters for 16 GRs for this purpose. Each consists of a single-copy plasmid coding for GFP fused to the full-length copy of the native promoter. We tracked their activity in exponential and stationary growth, as well as under weak and strong stresses. We show that the reporters have high sensitivity and specificity to all stresses tested and detect single-cell variability in transcription rates. Given the influence of GRs on the TFN, we expect that the new library will contribute to dissecting global transcriptional stress-response programs ofE. coli. Moreover, the library can be invaluable in bioindustrial applications that tune those programs to, instead of cell growth, favor productivity while reducing energy consumption.Abstract Figure

Publisher

Cold Spring Harbor Laboratory

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