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 Chan, K.
Right arrow Articles by Falkow, S.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Chan, K.
Right arrow Articles by Falkow, S.

 Previous Article  |  Next Article 

Infection and Immunity, September 2005, p. 5438-5449, Vol. 73, No. 9
0019-9567/05/$08.00+0     doi:10.1128/IAI.73.9.5438-5449.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.

Microarray-Based Detection of Salmonella enterica Serovar Typhimurium Transposon Mutants That Cannot Survive in Macrophages and Mice

Kaman Chan,* Charles C. Kim,{dagger} and Stanley Falkow

Department of Microbiology and Immunology, Stanford University School of Medicine, 299 Campus Drive, Stanford, California 94305-5124

Received 11 March 2005/ Returned for modification 19 April 2005/ Accepted 5 May 2005

DNA microarrays provide an opportunity to combine the principles of signature-tagged mutagenesis (STM) with microarray technology to identify potentially important bacterial virulence genes. The scope of DNA microarrays allows for less laborious screening on a much larger scale than possible by STM alone. We have adapted a microarray-based transposon tracking strategy for use with a Salmonella enterica serovar Typhimurium cDNA microarray in order to identify genes important for survival and replication in RAW 264.7 mouse macrophage-like cells or in the spleens of BALB/cJ mice. A 50,000-CFU transposon library of S. enterica serovar Typhimurium strain SL1344 was serially passaged in cultured macrophages or intraperitoneally inoculated into BALB/cJ mice. The bacterial genomic DNA was isolated and processed for analysis on the microarray. The novel application of this approach to identify mutants unable to survive in cultured cells resulted in the identification of components of Salmonella pathogenicity island 2 (SPI2), which is known to be critical for intracellular survival and replication. In addition, array results indicated that a number of SPI1-associated genes, currently not associated with intracellular survival, are negatively selected. However, of the SPI1-associated mutants individually tested for intracellular survival, only a sirA mutant exhibited reduced numbers relative to those of wild-type bacteria. Of the mutants unable to survive in mice, significant proportions are either components of the SPI2 pathogenicity island or involved in lipopolysaccharide synthesis. This observation is in agreement with results obtained in the original S. enterica serovar Typhimurium STM screen, illustrating the utility of this approach for the high-throughput identification of virulence factors important for survival in the host.

* Corresponding author. Mailing address: Department of Microbiology and Immunology, Stanford University School of Medicine, 299 Campus Drive, Stanford, CA 94305-5124. Phone: (650) 723-2671. Fax: (650) 723-1837. E-mail: kchan47{at}gmail.com.

Editor: A. D. O'Brien

{dagger} Present address: University of California, San Francisco, 600 16th St., Box 2240, San Francisco, CA 94158.

Infection and Immunity, September 2005, p. 5438-5449, Vol. 73, No. 9
0019-9567/05/$08.00+0     doi:10.1128/IAI.73.9.5438-5449.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.



This article has been cited by other articles:

  • Stevens, M. P., Humphrey, T. J., Maskell, D. J. (2009). Molecular insights into farm animal and zoonotic Salmonella infections. Phil Trans R Soc B 364: 2709-2723 [Abstract] [Full Text]  
  • Day, J. B., Basavanna, U., Sharma, S. K. (2009). Development of a Cell Culture Method To Isolate and Enrich Salmonella enterica Serotype Enteritidis from Shell Eggs for Subsequent Detection by Real-Time PCR. Appl. Environ. Microbiol. 75: 5321-5327 [Abstract] [Full Text]  
  • Zemansky, J., Kline, B. C., Woodward, J. J., Leber, J. H., Marquis, H., Portnoy, D. A. (2009). Development of a mariner-Based Transposon and Identification of Listeria monocytogenes Determinants, Including the Peptidyl-Prolyl Isomerase PrsA2, That Contribute to Its Hemolytic Phenotype. J. Bacteriol. 191: 3950-3964 [Abstract] [Full Text]  
  • Karsi, A., Gulsoy, N., Corb, E., Dumpala, P. R., Lawrence, M. L. (2009). High-Throughput Bioluminescence-Based Mutant Screening Strategy for Identification of Bacterial Virulence Genes. Appl. Environ. Microbiol. 75: 2166-2175 [Abstract] [Full Text]  
  • Jackson, L. K., Nawabi, P., Hentea, C., Roark, E. A., Haldar, K. (2008). The Salmonella virulence protein SifA is a G protein antagonist. Proc. Natl. Acad. Sci. USA 105: 14141-14146 [Abstract] [Full Text]  
  • Menashe, O., Kaganskaya, E., Baasov, T., Yaron, S. (2008). Aminoglycosides Affect Intracellular Salmonella enterica Serovars Typhimurium and Virchow. Antimicrob. Agents Chemother. 52: 920-926 [Abstract] [Full Text]  
  • Huang, Y., Leming, C. L., Suyemoto, M., Altier, C. (2007). Genome-Wide Screen of Salmonella Genes Expressed during Infection in Pigs, Using In Vivo Expression Technology. Appl. Environ. Microbiol. 73: 7522-7530 [Abstract] [Full Text]  
  • Lane, M. C., Alteri, C. J., Smith, S. N., Mobley, H. L. T. (2007). Expression of flagella is coincident with uropathogenic Escherichia coli ascension to the upper urinary tract. Proc. Natl. Acad. Sci. USA 104: 16669-16674 [Abstract] [Full Text]  
  • Paulin, S. M., Jagannathan, A., Campbell, J., Wallis, T. S., Stevens, M. P. (2007). Net Replication of Salmonella enterica Serovars Typhimurium and Choleraesuis in Porcine Intestinal Mucosa and Nodes Is Associated with Their Differential Virulence. Infect. Immun. 75: 3950-3960 [Abstract] [Full Text]  
  • Fink, R. C., Evans, M. R., Porwollik, S., Vazquez-Torres, A., Jones-Carson, J., Troxell, B., Libby, S. J., McClelland, M., Hassan, H. M. (2007). FNR Is a Global Regulator of Virulence and Anaerobic Metabolism in Salmonella enterica Serovar Typhimurium (ATCC 14028s). J. Bacteriol. 189: 2262-2273 [Abstract] [Full Text]  
  • Bijlsma, J. J. E., Burghout, P., Kloosterman, T. G., Bootsma, H. J., de Jong, A., Hermans, P. W. M., Kuipers, O. P. (2007). Development of Genomic Array Footprinting for Identification of Conditionally Essential Genes in Streptococcus pneumoniae. Appl. Environ. Microbiol. 73: 1514-1524 [Abstract] [Full Text]  
  • Baldwin, D. N., Shepherd, B., Kraemer, P., Hall, M. K., Sycuro, L. K., Pinto-Santini, D. M., Salama, N. R. (2007). Identification of Helicobacter pylori Genes That Contribute to Stomach Colonization. Infect. Immun. 75: 1005-1016 [Abstract] [Full Text]  
  • Mondragon, V., Franco, B., Jonas, K., Suzuki, K., Romeo, T., Melefors, O., Georgellis, D. (2006). pH-Dependent Activation of the BarA-UvrY Two-Component System in Escherichia coli. J. Bacteriol. 188: 8303-8306 [Abstract] [Full Text]  
  • Deiwick, J., Salcedo, S. P., Boucrot, E., Gilliland, S. M., Henry, T., Petermann, N., Waterman, S. R., Gorvel, J.-P., Holden, D. W., Meresse, S. (2006). The Translocated Salmonella Effector Proteins SseF and SseG Interact and Are Required To Establish an Intracellular Replication Niche. Infect. Immun. 74: 6965-6972 [Abstract] [Full Text]  
  • Nagy, G., Danino, V., Dobrindt, U., Pallen, M., Chaudhuri, R., Emody, L., Hinton, J. C., Hacker, J. (2006). Down-Regulation of Key Virulence Factors Makes the Salmonella enterica Serovar Typhimurium rfaH Mutant a Promising Live-Attenuated Vaccine Candidate.. Infect. Immun. 74: 5914-5925 [Abstract] [Full Text]  
  • Terekhova, D., Iyer, R., Wormser, G. P., Schwartz, I. (2006). Comparative Genome Hybridization Reveals Substantial Variation among Clinical Isolates of Borrelia burgdorferi Sensu Stricto with Different Pathogenic Properties.. J. Bacteriol. 188: 6124-6134 [Abstract] [Full Text]  
  • Faucher, Séb. P., Porwollik, S., Dozois, C. M., McClelland, M., Daigle, F. (2006). Transcriptome of Salmonella enterica serovar Typhi within macrophages revealed through the selective capture of transcribed sequences. Proc. Natl. Acad. Sci. USA 103: 1906-1911 [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»