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Infection and Immunity, December 2001, p. 7736-7742, Vol. 69, No. 12
0019-9567/01/$04.00+0 DOI: 10.1128/IAI.69.12.7736-7742.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.
Brucella abortus Genes Identified
following Constitutive Growth and Macrophage Infection
Linda
Eskra,
Aurea
Canavessi,
Merriann
Carey, and
Gary
Splitter*
Department of Animal Health and Biomedical
Sciences, University of Wisconsin
Madison, Madison, Wisconsin
53706
Received 16 April 2001/Returned for modification 2 July
2001/Accepted 20 August 2001
The chronicity of Brucella abortus infection in
humans and animals depends on the organism's ability to escape host
defenses by gaining entry and surviving inside the macrophage. Although no human vaccine exists for Brucella, vaccine
development in other bacteria has been based on deletions of selective
nutritional as well as regulatory systems. Our goal is to develop a
vaccine for Brucella. To further this aim, we have used
a green fluorescent protein (GFP) reporter system to identify
constitutively and intracellularly induced B. abortus
genes. Constitutively producing gfp clones exhibited
sequence homology with genes associated with protein synthesis and
metabolism (initiation factor-1 and tRNA ribotransferase) and
detoxification (organic hydroperoxidase resistance). Of greater interest, clones negative for constitutively produced
gfp in agar were examined by fluorescence microscopy to
detect promoter activity induced within macrophages 4 and 24 h
following infection. Bacterial genes activated in macrophages 4 h
postinfection appear to be involved in adapting to intracellular
environmental conditions. Included in this group were genes for
detoxification (lactoglyglutathione lyase gene), repair
(formamidopyrimidine-DNA glycosylase gene), osmotic protection
(K+ transport gene), and site-specific recombination
(xerD gene). A gene involved in metabolism and
biosynthesis (deoxyxylulose 5' phosphate synthase gene) was also
identified. Genes activated 24 h following infection were
biosynthesis- and metabolism-associated genes (iron binding protein and
rhizopine catabolism). Identification of B. abortus
genes that are activated following macrophage invasion provides insight
into Brucella pathogenesis and thus is valuable in
vaccine design utilizing selective targeted deletions of newly identified Brucella genes.
*
Corresponding author. Mailing address: Department of
Animal Health and Biomedical Sciences, University of
Wisconsin-Madison, 1656 Linden Dr., Madison, WI 53706. Phone: (608)
262-1837. Fax: (608) 262-7420. E-mail:
Splitter{at}ahabs.wisc.edu.
Infection and Immunity, December 2001, p. 7736-7742, Vol. 69, No. 12
0019-9567/01/$04.00+0 DOI: 10.1128/IAI.69.12.7736-7742.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.
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