Subcellular localization of legionella Dot/Icm effectors

Adam J Vogrin, Aurelie Mousnier, Gad Frankel, Elizabeth L Hartland

Research output: Contribution to journalArticle

Abstract

The translocation of effector proteins by the Dot/Icm type IV secretion system is central to the ability of Legionella pneumophila to persist and replicate within eukaryotic cells. The subcellular localization of translocated Dot/Icm proteins in host cells provides insight into their function. Through co-staining with host cell markers, effector proteins may be localized to specific subcellular compartments and membranes, which frequently reflects their host cell target and mechanism of action. In this chapter, we describe protocols to (1) localize effector proteins within cells by ectopic expression using green fluorescent protein fusions and (2) localize effector proteins within infected cells using epitope-tagged effector proteins and immuno-fluorescence microscopy.

Original languageEnglish
Pages (from-to)333-44
Number of pages12
JournalMethods in Molecular Biology
Volume954
DOIs
Publication statusPublished - 2013

Keywords

  • Bacterial Proteins
  • Bacterial Secretion Systems
  • Cloning, Molecular
  • Gene Expression
  • Green Fluorescent Proteins
  • HEK293 Cells
  • Humans
  • Legionella
  • Legionella pneumophila
  • Microscopy, Fluorescence
  • Plasmids
  • Protein Transport
  • Recombinant Fusion Proteins
  • Transfection

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