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Streptococcus pneumoniae PstS production is phosphate responsive and enhanced during growth in the murine peritoneal cavityDifferential display-PCR (DDPCR) was used to identify a Streptococcus pneumoniae gene with enhanced transcription during growth in the murine peritoneal cavity. Northern dot blot analysis and comparative densitometry confirmed a 1.8-fold increase in expression of the encoded sequence following murine peritoneal culture (MPC) versus laboratory culture or control culture (CC). Sequencing and basic local alignment search tool analysis identified the DDPCR fragment as pstS, the phosphate-binding protein of a high-affinity phosphate uptake system. PCR amplification of the complete pstS gene followed by restriction analysis and sequencing suggests a high level of conservation between strains and serotypes. Quantitative immunodot blotting using antiserum to recombinant PstS (rPstS) demonstrated an approximately twofold increase in PstS production during MPC from that during CCs, a finding consistent with the low levels of phosphate observed in the peritoneum. Moreover, immunodot blot and Northern analysis demonstrated phosphate-dependent production of PstS in six of seven strains examined. These results identify pstS expression as responsive to the MPC environment and extracellular phosphate concentrations. Presently, it remains unclear if phosphate concentrations in vivo contribute to the regulation of pstS. Finally, polyclonal antiserum to rPstS did not inhibit growth of the pneumococcus in vitro, suggesting that antibodies do not block phosphate uptake; moreover, vaccination of mice with rPstS did not protect against intraperitoneal challenge as assessed by the 50% lethal dose.
Document ID
20040088663
Acquisition Source
Legacy CDMS
Document Type
Reprint (Version printed in journal)
Authors
Orihuela, C. J.
(The University of Texas Medical Branch at Galveston, Department of Microbiology and Immunology Galveston, Texas 77555-1019, United States)
Mills, J.
Robb, C. W.
Wilson, C. J.
Watson, D. A.
Niesel, D. W.
Date Acquired
August 21, 2013
Publication Date
December 1, 2001
Publication Information
Publication: Infection and immunity
Volume: 69
Issue: 12
ISSN: 0019-9567
Subject Category
Life Sciences (General)
Distribution Limits
Public
Copyright
Other
Keywords
Non-NASA Center
NASA Discipline Environmental Health

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