ArticleMicrobial biotechnology2020
Physical decoupling of XylS/Pm regulatory elements and conditional proteolysis enable precise control of gene expression in Pseudomonas putida.
Article in Microbial biotechnology, 2020. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 38 papers.
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Who cites it
38 citing papers in PubMed, 72 citations in OpenAlex.
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- Going beyond size: exploring the metabolic burden inSynthetic biology (Oxford, England) · 2026Article
- Cellulase secretion by engineered Pseudomonas putida enables growth on cellulose oligomers.Applied microbiology and biotechnology · 2025Article
- Combinatorial engineering pinpoints shikimate pathway bottlenecks in para-aminobenzoic acid production in Pseudomonas putida.Journal of biological engineering · 2025Article
- Engineering a New SsrA-Based Degradation Tag (LAA-LAA) and a Bacterial Synthetic Oscillator.ACS synthetic biology · 2025Article
- Lignin-Degrading Enzymes and the Potential ofMicroorganisms · 2025Review
- Genetic Dissection of Cyclic di-GMP Signalling in Pseudomonas aeruginosa via Systematic Diguanylate Cyclase Disruption.Microbial biotechnology · 2025Article
- Modular plasmid design for autonomous multi-protein expression in Escherichia coli.Journal of biological engineering · 2025Article
- Leveraging EngineeredACS synthetic biology · 2025Article
- Biosensor-Guided Engineering of a Baeyer-Villiger Monooxygenase for Aliphatic Ester Production.Chembiochem : a European journal of chemical biology · 2025Article
- AND Logic Based on Suppressor tRNAs Enables Stringent Control of Sliding Base Editors inACS synthetic biology · 2024Article
- Impact of fleQ Deficiency on Resource Allocation and Heterologous Gene Expression in Pseudomonas putida Across Various Growth Media.Microbial biotechnology · 2024Article
- State-of-the-art in engineering small molecule biosensors and their applications in metabolic engineering.SLAS technology · 2024Review
- Core and auxiliary functions of one-carbon metabolism inmSystems · 2023Article
- SEVA 4.0: an update of the Standard European Vector Architecture database for advanced analysis and programming of bacterial phenotypes.Nucleic acids research · 2023Article
- Synthetic metabolism forRSC chemical biology · 2022Article
- Role of the CrcB transporter of Pseudomonas putida in the multi-level stress response elicited by mineral fluoride.Environmental microbiology · 2022Article
- Developing a CRISPR-assisted base-editing system for genome engineering of Pseudomonas chlororaphis.Microbial biotechnology · 2022Article
- Bacterial degrons in synthetic circuits.Open biology · 2022Review
Corrections and comments
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Authors and funding
4 authors at 1 institution in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
Most of the gene expression systems available for Gram-negative bacteria are afflicted by relatively high levels of basal (i.e. leaky) expression of the target gene(s). This occurrence affects the system dynamics, ultimately reducing the output and productivity of engineered pathways and synthetic circuits. In order to circumvent this problem, we have designed a novel expression system based on the well-known XylS/Pm transcriptional regulator/promoter pair from the soil bacterium Pseudomonas putida mt-2, in which the key functional elements are physically decoupled. By integrating the xylS gene into the chromosome of the platform strain KT2440, while placing the Pm promoter into a set of standard plasmid vectors, the inducibility of the system (i.e. the output difference between the induced and uninduced state) improved up to 170-fold. We further combined this modular system with an extra layer of post-translational control by means of conditional proteolysis. In this setup, the target gene is tagged with a synthetic motif dictating protein degradation. When the system features were characterized using the monomeric superfolder GFP as a model protein, the basal levels of fluorescence were brought down to zero (i.e. below the limit of detection). In all, these novel expression systems constitute an alternative tool to altogether suppress leaky gene expression, and they can be easily adapted to other vector formats and plugged-in into different Gram-negative bacterial species at the user's will.
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