Profiling bacterial kinase activity using a genetic circuit

Eric van der Helm, Rasmus Bech, Christina Eva Lehning, Ruben Vazquez-Uribe, Morten Otto Alexander Sommer

Research output: Working paperResearch

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Abstract

Phosphorylation is a post-translational modification that regulates the activity of several key proteins in bacteria and eukaryotes. Accordingly, a variety of tools has been developed to measure kinase activity. To couple phosphorylation to an in vivo fluorescent readout we used the Bacillus subtilis kinase PtkA, transmembrane activator TkmA and the repressor FatR to construct a genetic circuit in E. coli. By tuning the repressor and kinase expression level at the same time, we were able to show a 4.2-fold increase in signal upon kinase induction. We furthermore validated that the previously reported FatR Y45E mutation1 attenuates operator repression. This genetic circuit provides a starting point for computational protein design and a metagenomic library-screening tool.
Original languageEnglish
PublisherNovo Nordisk Foundation Center for Biosustainability
Number of pages1
DOIs
Publication statusPublished - 2017

Cite this

van der Helm, E., Bech, R., Lehning, C. E., Vazquez-Uribe, R., & Sommer, M. O. A. (2017). Profiling bacterial kinase activity using a genetic circuit. Novo Nordisk Foundation Center for Biosustainability. https://doi.org/10.17605/OSF.IO/AKG5X
van der Helm, Eric ; Bech, Rasmus ; Lehning, Christina Eva ; Vazquez-Uribe, Ruben ; Sommer, Morten Otto Alexander. / Profiling bacterial kinase activity using a genetic circuit. Novo Nordisk Foundation Center for Biosustainability, 2017.
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van der Helm, E, Bech, R, Lehning, CE, Vazquez-Uribe, R & Sommer, MOA 2017 'Profiling bacterial kinase activity using a genetic circuit' Novo Nordisk Foundation Center for Biosustainability. https://doi.org/10.17605/OSF.IO/AKG5X

Profiling bacterial kinase activity using a genetic circuit. / van der Helm, Eric; Bech, Rasmus; Lehning, Christina Eva; Vazquez-Uribe, Ruben; Sommer, Morten Otto Alexander.

Novo Nordisk Foundation Center for Biosustainability, 2017.

Research output: Working paperResearch

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van der Helm E, Bech R, Lehning CE, Vazquez-Uribe R, Sommer MOA. Profiling bacterial kinase activity using a genetic circuit. Novo Nordisk Foundation Center for Biosustainability. 2017. https://doi.org/10.17605/OSF.IO/AKG5X