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Infection and Immunity, March 2004, p. 1265-1274, Vol. 72, No. 3
0019-9567/04/$08.00+0     DOI: 10.1128/IAI.72.3.1265-1274.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.

A 38-Kilobase Pathogenicity Island Specific for Mycobacterium avium subsp. paratuberculosis Encodes Cell Surface Proteins Expressed in the Host

Janin Stratmann,^1, Birgit Strommenger,^1, Ralph Goethe,¹ Karen Dohmann,¹ Gerald-F. Gerlach,^1* Karen Stevenson,² Ling-ling Li,³ Qing Zhang,³ Vivek Kapur,³ and Tim J. Bull⁴

Institute for Microbiology, Department of Infectious Diseases, School of Veterinary Medicine, Hannover, Germany,¹ Moredun Research Institute, International Research Centre, Penicuik, Scotland,² Department of Surgery, St. George's Hospital Medical School, London, United Kingdom,⁴ Department of Microbiology and Biomedical Genomics Center, University of Minnesota, Minneapolis, Minnesota³

Received 12 June 2003/ Returned for modification 4 September 2003/ Accepted 13 November 2003

We have used representational difference analysis to identify a novel Mycobacterium avium subsp. paratuberculosis-specific ABC transporter operon (mpt), which comprises six open reading frames designated mptA to -F and is immediately preceded by two putative Fur boxes. Functional genomics revealed that the mpt operon is flanked on one end by a fep cluster encoding proteins involved in the uptake of Fe³⁺ and on the other end by a sid cluster encoding non-ribosome-dependent heterocyclic siderophore synthases. Together these genes form a 38-kb M. avium subsp. paratuberculosis-specific locus flanked by an insertion sequence similar to IS1110. Expression studies using Western blot analyses showed that MptC is present in the envelope fraction of M. avium subsp. paratuberculosis. The MptD protein was shown to be surface exposed, using a specific phage (fMptD) isolated from a phage-peptide library, by differential screening of Mycobacterium smegmatis transformants. The phage fMptD-derived peptide could be used in a peptide-mediated capture PCR with milk from infected dairy herds, thereby showing surface-exposed expression of the MptD protein in the host. Together, these data suggest that the 38-kb locus constitutes an M. avium subsp. paratuberculosis pathogenicity island.

Editor: B. B. Finlay

J.S. and B.S. contributed equally to this work.

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