A tunable anthranilate-inducible gene expression system for Pseudomonas species

Area of Science:

• Microbiology
• Molecular Biology
• Biotechnology

Background:

• Pseudomonads are ubiquitous bacteria found in diverse environments.
• Existing gene expression systems for pseudomonads lack tunability and broad applicability.
• There is a need for a simple, adaptable system for controlled gene expression in pseudomonads.

Purpose of the Study:

• To establish a tunable gene expression system for a wide range of pseudomonads.
• To develop an anthranilate-inducible promoter system (P_antA) for controlled gene expression.
• To assess the system's utility in physiological and biotechnological applications.

Main Methods:

• Construction of an anthranilate-inducible AntR/P_antA-based expression system (pUCP20-ANT) in *P. fluorescens*.
• Utilized Green Fluorescent Protein (GFP) as a reporter to analyze gene expression.
• Compared the P_antA system with a rhamnose-inducible RhaSR/P_rhaB system.

Main Results:

• The P_antA system demonstrated tunable gene expression across a range of anthranilate concentrations.
• Unlike the rhamnose system, P_antA allowed fine-tuning of expression levels, from low to high.
• The anthranilate-inducible system proved effective in *P. aeruginosa* and *P. putida* strains.

Conclusions:

• The anthranilate-inducible P_antA promoter provides a versatile tool for tunable gene expression in pseudomonads.
• This system offers significant advantages for physiological studies and biotechnological applications.
• The developed system is broadly applicable to various pseudomonad species.