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Science 26 September 1997:
Vol. 277. no. 5334, pp. 2007 - 2011
DOI: 10.1126/science.277.5334.2007
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Reports

Fluorescence-Based Isolation of Bacterial Genes Expressed Within Host Cells

Raphael H. Valdivia, * Stanley Falkow

A selection strategy was devised to identify bacterial genes preferentially expressed when a bacterium associates with its host cell. Fourteen Salmonella typhimurium genes, which were under the control of at least four independent regulatory circuits, were identified to be selectively induced in host macrophages. Four genes encode virulence factors, including a component of a type III secretory apparatus. This selection methodology should be generally applicable to the identification of genes from pathogenic organisms that are induced upon association with host cells or tissues.

Department of Microbiology and Immunology, Stanford University School of Medicine, Stanford, CA 94305, USA.
*   To whom correspondence should be addressed. E-mail: valdivia{at}cmgm.stanford.edu

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N. R. Benson, R. M.-Y. Wong, and M. McClelland (2000)
J. Bacteriol. 182, 3490-3497
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Essential Role for Cholesterol in Entry of Mycobacteria into Macrophages.
J. Gatfield and J. Pieters (2000)
Science 288, 1647-1651
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Granuloma-Specific Expression of Mycobacterium Virulence Proteins from the Glycine-Rich PE-PGRS Family.
L. Ramakrishnan, N. A. Federspiel, and S. Falkow (2000)
Science 288, 1436-1439
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Applications of Flow Cytometry to Clinical Microbiology.
A. Alvarez-Barrientos, J. Arroyo, R. Canton, C. Nombela, and M. Sanchez-Perez (2000)
Clin. Microbiol. Rev. 13, 167-195
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Multiple Factors Independently Regulate hilA and Invasion Gene Expression in Salmonella enterica Serovar Typhimurium.
R. L. Lucas, C. P. Lostroh, C. C. DiRusso, M. P. Spector, B. L. Wanner, and C. A. Lee (2000)
J. Bacteriol. 182, 1872-1882
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Salmonella Pathogenicity Island 2-Dependent Evasion of the Phagocyte NADPH Oxidase.
A. Vazquez-Torres, Y. Xu, J. Jones-Carson, D. W. Holden, S. M. Lucia, M. C. Dinauer, P. Mastroeni, and F. C. Fang (2000)
Science 287, 1655-1658
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Use of Lambda Phage S and R Gene Products in an Inducible Lysis System for Vibrio cholerae- and Salmonella enterica Serovar Typhimurium-Based DNA Vaccine Delivery Systems.
V. Jain and J. J. Mekalanos (2000)
Infect. Immun. 68, 986-989
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OmpR Regulates the Two-Component System SsrA-SsrB in Salmonella Pathogenicity Island 2.
A. K. Lee, C. S. Detweiler, and S. Falkow (2000)
J. Bacteriol. 182, 771-781
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Carbonic anhydrase is an ancient enzyme widespread in prokaryotes.
K. S. Smith, C. Jakubzick, T. S. Whittam, and J. G. Ferry (1999)
PNAS 96, 15184-15189
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Constitutive and Inducible Expression of Green Fluorescent Protein in Brucella suis.
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Infect. Immun. 67, 6695-6697
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Salmonella typhimurium Virulence Genes Are Induced upon Bacterial Invasion into Phagocytic and Nonphagocytic Cells.
C. G. Pfeifer, S. L. Marcus, O. Steele-Mortimer, L. A. Knodler, and B. B. Finlay (1999)
Infect. Immun. 67, 5690-5698
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Resistance to Bacitracin as Modulated by an Escherichia coli Homologue of the Bacitracin ABC Transporter BcrC Subunit from Bacillus licheniformis.
Y. M. Harel, A. Bailone, and E. Bibi (1999)
J. Bacteriol. 181, 6176-6178
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Use of fluorescence induction and sucrose counterselection to identify Mycobacterium tuberculosis genes expressed within host cells.
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