This Article
Right arrow Full Text
Right arrow Full Text (PDF)
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Services
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Right arrowReprints and Permissions
Right arrow Copyright Information
Right arrow Books from ASM Press
Right arrow MicrobeWorld
Citing Articles
Right arrow Citing Articles via HighWire
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Wilson, R. L.
Right arrow Articles by Harty, J. T.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Wilson, R. L.
Right arrow Articles by Harty, J. T.

 Previous Article  |  Next Article 

Infection and Immunity, August 2001, p. 5016-5024, Vol. 69, No. 8
0019-9567/01/$04.00+0   DOI: 10.1128/IAI.69.8.5016-5024.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.

Identification of Listeria monocytogenes In Vivo-Induced Genes by Fluorescence-Activated Cell Sorting

Rebecca L. Wilson,* A. R. Tvinnereim, Bradley D. Jones, and John T. Harty

Department of Microbiology, University of Iowa, Iowa City, Iowa 52242

Received 20 November 2000/Returned for modification 20 February 2001/Accepted 10 May 2001

Listeria monocytogenes is a gram-positive, intracellular, food-borne pathogen capable of causing severe infections in immunocompromised or pregnant individuals, as well as numerous animal species. Genetic analysis of Listeria pathogenesis has identified several genes which are crucial for virulence. The transcription of most of these genes has been shown to be induced upon entry of Listeria into the host cell. To identify additional genes that are induced in vivo and may be required for L. monocytogenes pathogenesis, a fluorescence-activated cell-sorting technique was initiated. Random fragments of the L. monocytogenes chromosome were cloned into a plasmid carrying a promoterless green fluorescent protein (GFP) gene, and the plasmids were transformed into the L. monocytogenes actA mutant DP-L1942. Fluorescence-activated cell sorting (FACS) was used to isolate L. monocytogenes clones that exhibited increased GFP expression within macrophage-like J774 cells but had relatively low levels of GFP expression when the bacteria were extracellular. Using this strategy, several genes were identified, including actA, that exhibited such an expression profile. In-frame deletions of two of these genes, one encoding the putative L. monocytogenes uracil DNA glycosylase (ung) and one encoding a protein with homology to the Bacillus subtilis YhdP hemolysin-like protein, were constructed and introduced into the chromosome of wild-type L. monocytogenes 10403s. The L. monocytogenes 10403s ung deletion mutant was not attenuated for virulence in mice, while the yhdP mutant exhibited a three- to sevenfold reduction in virulence.

* Corresponding author. Mailing address: Department of Microbiology, University of Iowa, Iowa City, IA 52242. Phone: (319) 335-7794. Fax: (319) 335-9006. E-mail: rlwilson{at}blue.weeg.uiowa.edu.

Infection and Immunity, August 2001, p. 5016-5024, Vol. 69, No. 8
0019-9567/01/$04.00+0   DOI: 10.1128/IAI.69.8.5016-5024.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.


This article has been cited by other articles:

  • Richards, M. I., Michell, S. L., Oyston, P. C. F. (2008). An intracellularly inducible gene involved in virulence and polyphosphate production in Francisella. J Med Microbiol 57: 1183-1192 [Abstract] [Full Text]  
  • Dubail, I., Bigot, A., Lazarevic, V., Soldo, B., Euphrasie, D., Dupuis, M., Charbit, A. (2006). Identification of an Essential Gene of Listeria monocytogenes Involved in Teichoic Acid Biogenesis.. J. Bacteriol. 188: 6580-6591 [Abstract] [Full Text]  
  • Chatterjee, S. S., Hossain, H., Otten, S., Kuenne, C., Kuchmina, K., Machata, S., Domann, E., Chakraborty, T., Hain, T. (2006). Intracellular Gene Expression Profile of Listeria monocytogenes. Infect. Immun. 74: 1323-1338 [Abstract] [Full Text]  
  • Palumbo, J. D., Kaneko, A., Nguyen, K. D., Gorski, L. (2005). Identification of Genes Induced in Listeria monocytogenes during Growth and Attachment to Cut Cabbage, Using Differential Display. Appl. Environ. Microbiol. 71: 5236-5243 [Abstract] [Full Text]  
  • Rediers, H., Rainey, P. B., Vanderleyden, J., De Mot, R. (2005). Unraveling the Secret Lives of Bacteria: Use of In Vivo Expression Technology and Differential Fluorescence Induction Promoter Traps as Tools for Exploring Niche-Specific Gene Expression. Microbiol. Mol. Biol. Rev. 69: 217-261 [Abstract] [Full Text]  
  • Hautefort, I., Proenca, M. J., Hinton, J. C. D. (2003). Single-Copy Green Fluorescent Protein Gene Fusions Allow Accurate Measurement of Salmonella Gene Expression In Vitro and during Infection of Mammalian Cells. Appl. Environ. Microbiol. 69: 7480-7491 [Abstract] [Full Text]  
  • Mason, K. M., Munson, R. S. Jr., Bakaletz, L. O. (2003). Nontypeable Haemophilus influenzae Gene Expression Induced In Vivo in a Chinchilla Model of Otitis Media. Infect. Immun. 71: 3454-3462 [Abstract] [Full Text]  
  • Runyen-Janecky, L. J., Payne, S. M. (2002). Identification of Chromosomal Shigella flexneri Genes Induced by the Eukaryotic Intracellular Environment. Infect. Immun. 70: 4379-4388 [Abstract] [Full Text]  


Copyright © 2009 by the American Society for Microbiology.   For an alternate route to Journals.ASM.org, visit: http://intl-journals.asm.org | More Info»