Synthetic vanillate-regulated promoter for graded gene expression in Sphingomonas
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Date
2014-09-29
Publication Type
Journal Article
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yes
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Abstract
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.
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published
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Journal / series
Volume
4
Pages / Article No.
6453
Publisher
Nature
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Subject
Bacterial techniques and applications; Bacterial genes
Organisational unit
03740 - Vorholt, Julia / Vorholt, Julia
Notes
Funding
152835 - Investigation of the alphaproteobacterial general stress response in the context of the phyllosphere environment (SNF)