Use of transcriptional control to increase secretion of heterologous proteins in T3S systems

Kevin J. Metcalf, Danielle Tullman-Ercek*

*Corresponding author for this work

Research output: Chapter in Book/Report/Conference proceedingChapter

1 Scopus citations

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.

Original languageEnglish (US)
Title of host publicationMethods in Molecular Biology
PublisherHumana Press Inc.
Pages71-79
Number of pages9
DOIs
StatePublished - Jan 1 2017

Publication series

NameMethods in Molecular Biology
Volume1531
ISSN (Print)1064-3745

Keywords

  • Biotechnology
  • Heterologous protein
  • Protein secretion
  • Synthetic biology
  • Type III secretion

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics

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