Microbiology
HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS
QUICK SEARCH:   [advanced]


     

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
Right arrow Citation Map
Services
Right arrow Email this article to a friend
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 arrow reprints & permissions
Citing Articles
Right arrow Citing Articles via HighWire
Right arrow Citing Articles via CrossRef
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Hobson, R. J.
Right arrow Articles by Dale, J. W.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Hobson, R. J.
Right arrow Articles by Dale, J. W.
Agricola
Right arrow Articles by Hobson, R. J.
Right arrow Articles by Dale, J. W.
Microbiology (2002), 148, 1571-1579.
© 2002 Society for General Microbiology

Research Paper

Use of an arrayed promoter-probe library for the identification of macrophage-regulated genes in Mycobacterium tuberculosis

Russell J. Hobsona,1, Alan J. A. McBrideb,1, Karen E. Kempsell2 and Jeremy W. Dale1

School of Biomedical and Life Sciences, University of Surrey, Guildford, Surrey GU2 7XH, UK1
GlaxoSmithKline Research and Development, Gunnels Wood Road, Stevenage, Hertfordshire SG1 2NY, UK2

Author for correspondence: Jeremy W. Dale. Tel: +44 1483 686484. Fax: +44 1483 300374. e-mail: j.dale{at}surrey.ac.uk

The survival of Mycobacterium tuberculosis within the human host after infection, especially within macrophages, is likely to require the activation of a number of mycobacterial genes. To identify such genes, a promoter-probe library was constructed in which fragments of M. tuberculosis H37Rv DNA were inserted upstream of a lacZ reporter gene, using an Escherichia coli–mycobacterial shuttle vector. Mycobacterium bovis Bacille Calmette–Guérin (BCG) was subsequently transformed with this library and 4800 BCG clones were arrayed in a 96-well microtitre format, enabling the testing of individual clones for promoter activity under a variety of conditions. From preliminary screening, 41 clones were selected that exhibited upregulation of lacZ expression when subjected to acidified sodium nitrite. Subsequent sequence analyses identified 26 of these clones as containing potential promoters. After measuring lacZ expression in BCG clones recovered from a THP-1 macrophage cell line, three genes were selected for assessment of their expression in M. tuberculosis during macrophage infection, by real-time RT-PCR. Two of these genes, Rv1265 (with unknown function) and Rv2711 (encoding the iron-dependent repressor protein IdeR), showed evidence of being upregulated within macrophages.

Keywords: gene expression, virulence, promoters

Abbreviations: BCG, Bacille Calmette–Guérin

a Present address: URC Neuroendocrinology, Bristol Royal Infirmary, Marlborough Street, Bristol BS1 8HW, UK.

b Present address: Microbiology and Immunobiology Department, School of Medicine, Queens University of Belfast, Grosvenor Road, Belfast BT12 6BN, Ireland.




This article has been cited by other articles:


Home page
 Infect. Immun.Home page
P. Fontan, V. Aris, S. Ghanny, P. Soteropoulos, and I. Smith
Global Transcriptional Profile of Mycobacterium tuberculosis during THP-1 Human Macrophage Infection
Infect. Immun., February 1, 2008; 76(2): 717 - 725.
[Abstract] [Full Text] [PDF]

Home page
 MicrobiologyHome page
V. Srivastava, C. Rouanet, R. Srivastava, B. Ramalingam, C. Locht, and B. S. Srivastava
Macrophage-specific Mycobacterium tuberculosis genes: identification by green fluorescent protein and kanamycin resistance selection
Microbiology, March 1, 2007; 153(3): 659 - 666.
[Abstract] [Full Text] [PDF]

Home page
 J. Bacteriol.Home page
G. Bai, L. A. McCue, and K. A. McDonough
Characterization of Mycobacterium tuberculosis Rv3676 (CRPMt), a Cyclic AMP Receptor Protein-Like DNA Binding Protein
J. Bacteriol., November 15, 2005; 187(22): 7795 - 7804.
[Abstract] [Full Text] [PDF]

Home page
 J. Bacteriol.Home page
H. Yesilkaya, J. W. Dale, N. J. C. Strachan, and K. J. Forbes
Natural Transposon Mutagenesis of Clinical Isolates of Mycobacterium tuberculosis: How Many Genes Does a Pathogen Need?
J. Bacteriol., October 1, 2005; 187(19): 6726 - 6732.
[Abstract] [Full Text] [PDF]

Home page
 Infect. Immun.Home page
Y. C. Manabe, C. L. Hatem, A. K. Kesavan, J. Durack, and J. R. Murphy
Both Corynebacterium diphtheriae DtxR(E175K) and Mycobacterium tuberculosis IdeR(D177K) Are Dominant Positive Repressors of IdeR-Regulated Genes in M. tuberculosis
Infect. Immun., September 1, 2005; 73(9): 5988 - 5994.
[Abstract] [Full Text] [PDF]

Home page
 J. Bacteriol.Home page
M. A. Gazdik and K. A. McDonough
Identification of Cyclic AMP-Regulated Genes in Mycobacterium tuberculosis Complex Bacteria under Low-Oxygen Conditions
J. Bacteriol., April 15, 2005; 187(8): 2681 - 2692.
[Abstract] [Full Text] [PDF]

Home page
 Clin. Microbiol. Rev.Home page
I. Smith
Mycobacterium tuberculosis Pathogenesis and Molecular Determinants of Virulence
Clin. Microbiol. Rev., July 1, 2003; 16(3): 463 - 496.
[Abstract] [Full Text] [PDF]


HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS
INT J SYST EVOL MICROBIOL MICROBIOLOGY J GEN VIROL
J MED MICROBIOL ALL SGM JOURNALS
Copyright © 2002 Society for General Microbiology.