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Related Experiment Video

Updated: Mar 3, 2026

Implementation of a Permeable Membrane Insert-based Infection System to Study the Effects of Secreted Bacterial Toxins on Mammalian Host Cells
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The EXIT Strategy: an Approach for Identifying Bacterial Proteins Exported during Host Infection.

E F Perkowski1, K E Zulauf1, D Weerakoon1

  • 1Department of Microbiology and Immunology, University of North Carolina-Chapel Hill, Chapel Hill, North Carolina, USA.

Mbio
|April 27, 2017
PubMed
Summary

Researchers developed a new genome-wide method, EXIT (exported technology), to identify bacterial exported proteins during infection. This technology successfully identified 593 proteins in Mycobacterium tuberculosis during murine infection, including those preferentially exported in vivo.

Keywords:
EXITMycobacterium tuberculosisbeta-lactamase reporterin vivomembrane proteinsprotein exportprotein secretionvirulence

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Area of Science:

  • Microbiology and Immunology
  • Bacterial Pathogenesis
  • Proteomics

Background:

  • Exported bacterial proteins are crucial for physiology and virulence, and are targets for antibiotics and immune responses.
  • Previous methods for identifying exported proteins relied on in vitro conditions, potentially missing proteins exported specifically during host infection.
  • Mycobacterium tuberculosis, the causative agent of tuberculosis, possesses numerous exported proteins involved in its pathogenesis.

Purpose of the Study:

  • To develop a genome-wide method for identifying bacterial proteins exported during host infection.
  • To apply this method to Mycobacterium tuberculosis during a murine infection model.
  • To identify novel virulence factors and drug targets by characterizing in vivo-exported proteins.

Main Methods:

  • Development of a novel genome-wide technology named EXIT (exported technology).
  • Application of EXIT to Mycobacterium tuberculosis during a mouse model of infection.
  • Comparative analysis of protein export under in vitro and in vivo conditions.

Main Results:

  • Successfully identified 593 exported proteins of Mycobacterium tuberculosis during murine infection.
  • Discovered 112 in vivo-induced exported proteins, showing significantly higher export during infection than in vitro.
  • Generated a comprehensive dataset for mapping the topology of M. tuberculosis membrane proteins.

Conclusions:

  • The EXIT technology is a powerful tool for identifying bacterial exported proteins on an unprecedented scale, particularly during infection.
  • Identification of in vivo-induced exported proteins provides critical insights into bacterial pathogenesis and host-pathogen interactions.
  • EXIT has broad applicability for studying exported proteins in other pathogens and experimental settings, aiding in the development of new diagnostics and therapeutics.