Analyzing Association of the Endoplasmic Reticulum with the Legionella pneumophilaContaining Vacuole

A unique feature of the intracellular life cycle of Legionella pneumophila is the interaction between the vacuole in which L. pneumophila resides and the endoplasmic reticulum of the host cell. This interaction is crucial for L. pneumophila to establish a niche in which the bacteria can replicate intracellularly. Microscopic analysis of endoplasmic reticulum (ER) markers during infection yields information regarding the nature of the recruited vesicles as well as the kinetics of their recruitment. The recruitment of YFP-KDEL, GFP-p58, calnexin, and myc-Sec22b to the L. pneumophila – containing vacuole can be assessed by fluorescence microscopy. Methods for detection of these various ER markers during infection of mammalian cells by L. pneumophila are described.