title: Use of Transcriptional Control to Increase Secretion of Heterologous Proteins in T3S Systems.
abstract: Heterologous proteins can be produced in a bacterial host and purified from the cellular constituents. Secretion of the protein of interest to the extracellular space simplifies the purification process and is thought to alleviate toxicity problems associated with intracellular accumulation of the protein of interest. In this protocol, we describe a strategy to engineer protein secretion in a bacterial culture using transcriptional control. The transcription factor HilA is inducibly produced to control production of the secretion machine, and in turn signals the production and secretion of a protein of interest. This allows for high titer of secreted protein in optimized culturing conditions and the effect is observed with all proteins tested.
authors.affiliation: Department of Chemical and Biomolecular Engineering, University of California Berkeley, 116 Gilman Hall, Berkeley, CA, 94720, USA.
authors.forename: Kevin J
authors.initials: KJ
authors.lastname: Metcalf
authors.affiliation: Department of Chemical and Biomolecular Engineering, University of California Berkeley, 116 Gilman Hall, Berkeley, CA, 94720, USA. ercek@northwestern.edu.
authors.forename: Danielle
authors.initials: D
authors.lastname: Tullman-Ercek
mesh_terms: D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism; D015153:Blotting, Western; D015964:Gene Expression Regulation, Bacterial; D006090:Gram-Negative Bacteria / Q000235:genetics* / Q000378:metabolism*; D015534:Trans-Activators / Q000378:metabolism; D014158:Transcription, Genetic; D000069296:Type III Secretion Systems / Q000235:genetics* / Q000378:metabolism*
chemical_list: D001426:Bacterial Proteins; D015534:Trans-Activators; D000069296:Type III Secretion Systems
keywords: Biotechnology; Heterologous protein; Protein secretion; Synthetic biology; Type III secretion