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Dictyostelium Host Response to Legionella Infection: Strategies and Assays.
MedLine Citation:
PMID:  23150412     Owner:  NLM     Status:  Publisher    
Abstract/OtherAbstract:
The professional phagocyte Dictyostelium discoideum is a simple eukaryotic microorganism, whose natural habitat is deciduous forest soil and decaying leaves, where the amoebae feed on bacteria and grow as separate, independent, single cells. In the last decade, the organism has been successfully used as a host for several human pathogens, including Legionella pneumophila, Mycobacterium avium and Mycobacterium marinum,Pseudomonas aeruginosa, Klebsiella pneumoniae, Cryptococcus neoformans, and Salmonella typhimurium. To dissect the complex cross-talk between host and pathogen Dictyostelium offers easy cultivation, a high quality genome sequence and excellent molecular genetic and biochemical tools. Dictyostelium cells are also extremely suitable for cell biological studies, which in combination with in vivo expression of fluorescence-tagged proteins allow investigating the dynamics of bacterial uptake and infection. Inactivation of genes by homologous recombination as well as gene rescue and overexpression are well established and a large mutant collection is available at the Dictyostelium stock center, favoring identification of host resistance or susceptibility genes. Here, we briefly introduce the organism, address the value of Dictyostelium as model host, describe strategies to identify host cell factors important for infection followed by protocols for cell culture and storage, uptake and infection, and confocal microscopy of infected cells.
Authors:
Salvatore Bozzaro; Barbara Peracino; Ludwig Eichinger
Publication Detail:
Type:  JOURNAL ARTICLE    
Journal Detail:
Title:  Methods in molecular biology (Clifton, N.J.)     Volume:  954     ISSN:  1940-6029     ISO Abbreviation:  Methods Mol. Biol.     Publication Date:  2013  
Date Detail:
Created Date:  2012-11-14     Completed Date:  -     Revised Date:  -    
Medline Journal Info:
Nlm Unique ID:  9214969     Medline TA:  Methods Mol Biol     Country:  -    
Other Details:
Languages:  ENG     Pagination:  417-438     Citation Subset:  -    
Affiliation:
Department of Clinical and Biological Sciences, University of Turin, Orbassano, Italy, salvatore.bozzaro@unito.it.
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