caa a22 4500 528786288 20181217030314.0 cr unu---uuuuu 180924e20140929xx s 000 0 eng 10.3929/ethz-b-000090618 doi 10.1038/srep06453 doi (ETHRESEARCH)oai:www.research-collection.ethz.ch:20.500.11850/90618 Kaczmarczyk Andreas Synthetic vanillate-regulated promoter for graded gene expression in Sphingomonas [Elektronische Daten] [Andreas Kaczmarczyk, Julia A. Vorholt, Anne Francez-Charlot] Sci Rep Open access ethresearch Regulated promoters are an important basic genetic tool allowing, for example, gene-dosage and gene depletion studies. We have previously described a cumate-inducible promoter (PQ5) that is functional in diverse Alphaproteobacteria. This promoter has been engineered by combining a synthetic minimal promoter, Psyn2, and operator sites and the repressor of the Pseudomonas putida F1 cym/cmt system. In the present study, we engineered a vanillate-regulated promoter using Psyn2 and the regulatory elements of the Caulobacter crescentus vanR-vanAB system. We show that the resulting promoter, which we called PV10, responds rapidly to the inducer vanillate with an induction ratio of about two orders of magnitude in Sphingomonas melonis Fr1. In contrast to the switch-like behavior of PQ5, PV10 shows a linear dose-response curve at intermediate vanillate concentrations, allowing graded gene expression. PV10 is functionally compatible with and independent of PQ5 and cumate, and vice versa, suggesting that both systems can be used simultaneously. Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International http://creativecommons.org/licenses/by-nc-nd/4.0 ethresearch Bacterial techniques and applications ethresearch Bacterial genes ethresearch Vorholt Julia A. joint author Francez-Charlot Anne joint author Scientific Reports London : Nature Publishing Group 4, p. 6453 2045-2322 http://hdl.handle.net/20.500.11850/90618 text/html WWW-Backlink auf das Repository (Open access) 1 Journal Article ethresearch ETHRESEARCH 856 40 http://hdl.handle.net/20.500.11850/90618 text/html WWW-Backlink auf das Repository (Open access) ETHRESEARCH 100 1- Kaczmarczyk Andreas ETHRESEARCH 700 1- Vorholt Julia A. joint author ETHRESEARCH 700 1- Francez-Charlot Anne joint author ETHRESEARCH 773 0- Scientific Reports London : Nature Publishing Group 4, p. 6453 2045-2322 BK010053 XK010053 XK010000 ETHRESEARCH ETHRESEARCH ETHRESEARCH Journal Article Open access