Generation of the Brucella melitensis ORFeome Version 1.1

  1. Amélie Dricot1,
  2. Jean-François Rual1,2,
  3. Philippe Lamesch1,2,
  4. Nicolas Bertin2,
  5. Denis Dupuy2,
  6. Tong Hao2,
  7. Christophe Lambert1,
  8. Régis Hallez1,
  9. Jean-Marc Delroisse1,
  10. Jean Vandenhaute1,
  11. Ignacio Lopez-Goñi3,
  12. Ignacio Moriyon3,
  13. Juan M. Garcia-Lobo4,
  14. Félix J. Sangari4,
  15. Alastair P. MacMillan5,
  16. Sally J. Cutler5,
  17. Adrian M. Whatmore5,
  18. Stephanie Bozak6,
  19. Reynaldo Sequerra6,
  20. Lynn Doucette-Stamm6,
  21. Marc Vidal2,
  22. David E. Hill2,
  23. Jean-Jacques Letesson1, and
  24. Xavier De Bolle1,7
  1. 1 Research Unit in Molecular Biology (URBM), University of Namur, 5000 Namur, Belgium
  2. 2 Center for Cancer Systems Biology and Department of Cancer Biology, Dana-Farber Cancer Institute and Department of Genetics, Harvard Medical School, Boston, Massachusetts 02115, USA
  3. 3 Departamento de Microbiologia, Universidad de Navarra, Pamplona 31008, Spain
  4. 4 Departamento de Biología Molecular, Universidad de Cantabria, Santander 39011, Spain
  5. 5 Department of Statutory and Exotic Bacterial Diseases, FAO/WHO Collaborating Centre for Reference and Research on Brucellosis, Veterinary Laboratories Agency, Weybridge, Surrey KT15 3NB, United Kingdom
  6. 6 Agencourt Biosciences Corporation, Beverly, Massachusetts 01915, USA

Abstract

The bacteria of the Brucella genus are responsible for a worldwide zoonosis called brucellosis. They belong to the α-proteobacteria group, as many other bacteria that live in close association with a eukaryotic host. Importantly, the Brucellae are mainly intracellular pathogens, and the molecular mechanisms of their virulence are still poorly understood. Using the complete genome sequence of Brucella melitensis, we generated a database of protein-coding open reading frames (ORFs) and constructed an ORFeome library of 3091 Gateway Entry clones, each containing a defined ORF. This first version of the Brucella ORFeome (v1.1) provides the coding sequences in a user-friendly format amenable to high-throughput functional genomic and proteomic experiments, as the ORFs are conveniently transferable from the Entry clones to various Expression vectors by recombinational cloning. The cloning of the Brucella ORFeome v1.1 should help to provide a better understanding of the molecular mechanisms of virulence, including the identification of bacterial protein-protein interactions, but also interactions between bacterial effectors and their host's targets.

Footnotes

  • [The following individuals kindly provided reagents, samples, or unpublished information as indicated in the paper: C. Baldwin and R. Goenka. The Open Biosystems company will act as a distributor of the Brucella ORFeome clones.]

  • Article and publication are at http://www.genome.org/cgi/doi/10.1101/gr.2456204.

  • ↵7 Corresponding author. E-MAIL xavier.debolle{at}fundp.ac.be; FAX 32-81-72-42-97.

    • Accepted April 28, 2004.
    • Received February 13, 2004.
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  1. doi: 10.1101/gr.2456204 Genome Res. 14: 2201-2206 Cold Spring Harbor Laboratory Press

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