Predictable, Tunable Protein Production in <i>Salmonella</i> for Studying Host-Pathogen Interactions.

Cooper, Kendal G; Chong, Audrey; Starr, Tregei; Finn, Ciaran E; Steele-Mortimer, Olivia

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

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

Afiliación

Cooper KG; Laboratory of Bacteriology, Rocky Mountain Laboratories, National Institutes of Allergy and Infectious Diseases, National Institutes of Health, Hamilton, MT, United States.
Chong A; Laboratory of Bacteriology, Rocky Mountain Laboratories, National Institutes of Allergy and Infectious Diseases, National Institutes of Health, Hamilton, MT, United States.
Starr T; Laboratory of Bacteriology, Rocky Mountain Laboratories, National Institutes of Allergy and Infectious Diseases, National Institutes of Health, Hamilton, MT, United States.
Finn CE; Laboratory of Bacteriology, Rocky Mountain Laboratories, National Institutes of Allergy and Infectious Diseases, National Institutes of Health, Hamilton, MT, United States.
Steele-Mortimer O; Laboratory of Bacteriology, Rocky Mountain Laboratories, National Institutes of Allergy and Infectious Diseases, National Institutes of Health, Hamilton, MT, United States.

Front Cell Infect Microbiol ; 7: 475, 2017.

Article en En | MEDLINE | ID: mdl-29201859

RESUMEN

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.

Asunto(s)

Genes Bacterianos/genética; Interacciones Huésped-Patógeno/genética; Plásmidos/genética; Regiones Promotoras Genéticas/genética; Salmonella/genética; Salmonella/metabolismo; Proteínas Bacterianas/genética; Citosol; Variaciones en el Número de Copia de ADN; Proteínas Fluorescentes Verdes; Células HeLa; Humanos; Salmonella/patogenicidad; Infecciones por Salmonella/genética; Infecciones por Salmonella/metabolismo; Salmonella enterica; Transactivadores/genética; Sistemas de Secreción Tipo III/genética

Palabras clave

Salmonella; fluorescent protein; intracellular; plasmid; promoter; synthetic biology

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Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Plásmidos / Salmonella / Regiones Promotoras Genéticas / Interacciones Huésped-Patógeno / Genes Bacterianos Tipo de estudio: Prognostic_studies / Risk_factors_studies Límite: Humans Idioma: En Revista: Front Cell Infect Microbiol Año: 2017 Tipo del documento: Article País de afiliación: Estados Unidos Pais de publicación: Suiza

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