A dual-reporter system for investigating and optimizing protein translation and folding in E. coli

Ariane Zutz, Louise Hamborg, Lasse Ebdrup Pedersen, Maher M. Kassem, Elena Papaleo, Anna Koza, Markus J. Herrgård, Sheila Ingemann Jensen, Kaare Teilum, Kresten Lindorff-Larsen, Alex Toftgaard Nielsen

Research output: Contribution to journalJournal articleResearchpeer-review

22 Downloads (Pure)


Strategies for investigating and optimizing the expression and folding of proteins for biotechnological and pharmaceutical purposes are in high demand. Here, we describe a dual-reporter biosensor system that simultaneously assesses in vivo protein translation and protein folding, thereby enabling rapid screening of mutant libraries. We have validated the dual-reporter system on five different proteins and find an excellent correlation between reporter signals and the levels of protein expression and solubility of the proteins. We further demonstrate the applicability of the dual-reporter system as a screening assay for deep mutational scanning experiments. The system enables high throughput selection of protein variants with high expression levels and altered protein stability. Next generation sequencing analysis of the resulting libraries of protein variants show a good correlation between computationally predicted and experimentally determined protein stabilities. We furthermore show that the mutational experimental data obtained using this system may be useful for protein structure calculations.
Original languageEnglish
Article number6093
JournalNature Communications
Number of pages15
Publication statusPublished - 2021


Dive into the research topics of 'A dual-reporter system for investigating and optimizing protein translation and folding in E. coli'. Together they form a unique fingerprint.

Cite this