Predictable, Tunable Protein Production in Salmonella for Studying Host-Pathogen Interactions

Area of Science:

• Synthetic biology
• Microbial genetics

Background:

• Episomal protein production in Salmonella is crucial for various applications.
• Controlling protein expression levels is challenging, often leading to toxicity or insufficient yields.

Purpose of the Study:

• To develop a system for predictable and tunable episomal protein production in Salmonella.
• To create a series of plasmids enabling fine-tuning of constitutive gene expression.

Main Methods:

• Utilized synthetic genetic elements, including variable-strength synthetic promoters and a synthetic transcriptional terminator.
• Incorporated plasmid copy number variation to modulate protein output.
• Developed a system for uniform production of fluorescent and endogenous proteins.

Main Results:

• Generated a series of plasmids enabling protein production over a wide dynamic range.
• Achieved uniform protein production across different expression levels.
• Demonstrated efficient titration of toxic protein effects through fine-tuned expression.

Conclusions:

• The developed synthetic genetic system enhances predictability and tunability of episomal protein production in Salmonella.
• This system provides a valuable tool for optimizing protein expression and managing toxicity in microbial systems.