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

oleh: Kendal G. Cooper, Audrey Chong, Tregei Starr, Ciaran E. Finn, Olivia Steele-Mortimer

Format: Article
Diterbitkan: Frontiers Media S.A. 2017-11-01

Deskripsi

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.