Examination of Salmonella gene expression in an infected mammalian host using the green fluorescent protein and two-colour flow cytometry

Molecular Microbiology
Dirk Bumann

Abstract

Quantitative data on Salmonella gene expression in infected hosts are largely lacking because of technical problems. One attractive reporter, the green fluorescent protein (GFP), is widely used in vitro but is difficult to quantify in infected tissues because of the preponderance of background particles with similar fluorescence. Here, bacterial GFP emission was spectrally distinguished from host autofluorescence by two-colour flow cytometry. Using this technique, the in vivo activity of three well-characterized promoters (PsicA, PssaH and PpagC) was determined. Their spatial and temporal activity patterns are in close agreement with predictions based on previous data and the colonization defects of corresponding deletion strains. To identify additional Salmonella promoters that are induced in infected animals, a genomic library was sorted by flow cytometry yielding four independent promoters. Genes expressed from PpibB and PsifA contribute to virulence, and chorismate mutase expressed from ParoQ might participate in aromatic acid biosynthesis, which is also required for virulence. Promoter P3g appears to be part of a mobile genetic element that is lacking in the completely sequenced strain LT2.

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Citations

Sep 22, 2012·Cancer Gene Therapy·M CroninM Tangney
Dec 6, 2011·Nucleic Acids Research·Sara LeschnerSiegfried Weiss
Feb 24, 2011·Infection and Immunity·Renée M TsolisAndreas J Bäumler
Feb 24, 2006·Infection and Immunity·Claudia Rollenhagen, Dirk Bumann
Oct 25, 2012·PLoS Pathogens·Somedutta BaratDirk Bumann
Jun 3, 2004·Proceedings of the National Academy of Sciences of the United States of America·Claudia RollenhagenDirk Bumann
Nov 21, 2002·Proceedings of the National Academy of Sciences of the United States of America·Michelle L ZaharikB Brett Finlay
Jan 21, 2016·Nature·Alexander J WestermannJörg Vogel
Sep 12, 2009·Current Opinion in Microbiology·Dirk Bumann
Mar 17, 2009·Current Opinion in Biotechnology·Helene L Andrews-PolymenisMichael McClelland
Oct 13, 2007·Journal of Microbiological Methods·Friederike ThöneDirk Bumann
Oct 5, 2006·Cellular Microbiology·Dominik E DorerAlexander D McLellan
Oct 20, 2010·FEMS Microbiology Reviews·Nuria AndreuSiouxsie Wiles
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Feb 1, 2005·FEMS Immunology and Medical Microbiology·Rita TamayoJohn S Gunn
Sep 27, 2018·PLoS Genetics·Lucía García-PastorJosep Casadesús
Apr 13, 2004·Applied Optics·Michael B SinclairMargaret Werner-Washburne

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