AUTHOR=Cooper Kendal G. , Chong Audrey , Starr Tregei , Finn Ciaran E. , Steele-Mortimer Olivia TITLE=Predictable, Tunable Protein Production in Salmonella for Studying Host-Pathogen Interactions JOURNAL=Frontiers in Cellular and Infection Microbiology VOLUME=7 YEAR=2017 URL=https://www.frontiersin.org/journals/cellular-and-infection-microbiology/articles/10.3389/fcimb.2017.00475 DOI=10.3389/fcimb.2017.00475 ISSN=2235-2988 ABSTRACT=

Here we describe the use of synthetic genetic elements to improve the predictability and tunability of episomal protein production in Salmonella. We used a multi-pronged approach, in which a series of variable-strength synthetic promoters were combined with a synthetic transcriptional terminator, and plasmid copy number variation. This yielded a series of plasmids that drive uniform production of fluorescent and endogenous proteins, over a wide dynamic range. We describe several examples where this system is used to fine-tune constitutive expression in Salmonella, providing an efficient means to titrate out toxic effects of protein production.