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Refactoring the Salmonella Type III Secretion System

  • Author(s): Mirsky, Ethan
  • Advisor(s): Voigt, Christopher
  • et al.
Abstract

Bacteria naturally contain a wide variety of multi-component molecular machines that perform complex functions such as chemotaxis, photosynthesis, chemical fixation, and protein secretion, all of which are potentially valuable to synthetic biologists for use in engineered organisms. However, in most cases, the gene clusters encoding these functions are complex and intricately regulated by the bacteria, making them difficult to reuse for new applications.

One potential method to address these difficulties is refactoring: a process of recoding the genes for the purpose of simplification and re-use. Here, we present the methods, techniques and results of refactoring the Salmonella Pathogenicity Island I (SPI-1) Type III Secretion System (T3SS). The SPI-1 T3SS is used naturally by Salmonella to inject pathogenic effector agents into its host cell and, unlike the most commonly used secretion mechanisms in bacteria, is capable of transporting functional proteins through both membranes of the bacteria directly into the culture media. This function may enable such biotech applications as targeted peptide/drug delivery and continuous protein expression. In addition, the methods developed here should be broadly applicable to a wide range of bacterial gene-clusters.

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