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Journal of Bacteriology, September 2009, p. 5377-5386, Vol. 191, No. 17
0021-9193/09/$08.00+0     doi:10.1128/JB.00597-09
Copyright © 2009, American Society for Microbiology. All Rights Reserved.

Proteomic and Genomic Characterization of Highly Infectious Clostridium difficile 630 Spores{triangledown} ,{dagger}

Trevor D. Lawley,1* Nicholas J. Croucher,2 Lu Yu,3 Simon Clare,1 Mohammed Sebaihia,2 David Goulding,1 Derek J. Pickard,1 Julian Parkhill,2 Jyoti Choudhary,3 and Gordon Dougan1

Microbial Pathogenesis Laboratory,1 Pathogen Genomics,2 Proteomics Mass Spectrometry Laboratory, The Wellcome Trust Sanger Institute, Wellcome Trust Genome Campus, Hinxton, CB10 1SA, United Kingdom3

Received 7 May 2009/ Accepted 12 June 2009

Clostridium difficile, a major cause of antibiotic-associated diarrhea, produces highly resistant spores that contaminate hospital environments and facilitate efficient disease transmission. We purified C. difficile spores using a novel method and show that they exhibit significant resistance to harsh physical or chemical treatments and are also highly infectious, with <7 environmental spores per cm2 reproducibly establishing a persistent infection in exposed mice. Mass spectrometric analysis identified ~336 spore-associated polypeptides, with a significant proportion linked to translation, sporulation/germination, and protein stabilization/degradation. In addition, proteins from several distinct metabolic pathways associated with energy production were identified. Comparison of the C. difficile spore proteome to those of other clostridial species defined 88 proteins as the clostridial spore "core" and 29 proteins as C. difficile spore specific, including proteins that could contribute to spore-host interactions. Thus, our results provide the first molecular definition of C. difficile spores, opening up new opportunities for the development of diagnostic and therapeutic approaches.

* Corresponding author. Mailing address: Microbial Pathogenesis Laboratory, Wellcome Trust Sanger Institute, Hinxton, Cambridgeshire, United Kingdom, CB10 1SA. Phone: 01223 495 391. Fax: 01223 495 239. E-mail: tl2{at}sanger.ac.uk

{triangledown} Published ahead of print on 19 June 2009.

{dagger} Supplemental material for this article may be found at http://jb.asm.org/.

Journal of Bacteriology, September 2009, p. 5377-5386, Vol. 191, No. 17
0021-9193/09/$08.00+0     doi:10.1128/JB.00597-09
Copyright © 2009, American Society for Microbiology. All Rights Reserved.





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