Isolation of the Dot/Icm Type IV Secretion System Core Complex from Legionella pneumophila
Legionella utilizes specialized protein secretion machinery called the type IV secretion system encoded by dot/icm genes to modulate host cellular systems. We describe here the procedure to isolate the core complex of the Dot/Icm type IV secretion system of L. pneumophila based on detergent lysis of bacteria and ultracentrifugation. The isolated protein complex can be applied for biochemical and transmission electron microscopy analysis.
Key wordsLegionella Type IV secretion Core complex Electron microscopy Nanomachine Molecular structure Ultracentrifugation
This work has been financially supported by MEXT/JSPS KAKENHI Grants 15H01322 (to TK). The protocol presented here has been adapted from Kubori et al. .
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