Development and use of a partial Mycobacterium avium subspecies paratuberculosis protein array

John P. Bannantine, W. Ray Waters, Judith R. Stabel, Mitchell V. Palmer, Lingling Li, Vivek Kapur, Michael L. Paustian

Research output: Contribution to journalArticle

16 Scopus citations

Abstract

As an initial step toward systematically characterizing all antigenic proteins produced by a significant veterinary pathogen, 43 recombinant Mycobacterium avium subspecies paratuberculosis (M. paratuberculosis) expression clones were constructed, cataloged, and stored. NC filters were spotted with purified proteins from each clone along with a whole cell lysate of M. paratuberculosis. Spots on the resulting dot array consisted of hypothetical proteins (13), metabolic proteins (3), cell envelope proteins (7), known antigens (4), and unique proteins with no similarity in public sequence databases (16). Dot blot arrays were used to profile antibody responses in a rabbit and mouse exposed to M. paratuberculosis as well as in cattle showing clinical signs of Johne's disease. The M. paratuberculosis heat shock protein DnaK, encoded by ORF MAP3840 and a membrane protein (MAP2121c), were identified as the most strongly immunoreactive in both the mouse and rabbit hosts, respectively. MAP3155c, which encodes a hypothetical protein, was most strongly immunoreactive in sera from Johne's disease cattle. This study has enabled direct comparisons of antibody reactivity for an entire panel of over 40 proteins and has laid the foundation for future high throughput production and arraying of M. paratuberculosis surface proteins for immune profiling experiments in cattle.

Original languageEnglish (US)
Pages (from-to)463-474
Number of pages12
JournalProteomics
Volume8
Issue number3
DOIs
StatePublished - Feb 1 2008

All Science Journal Classification (ASJC) codes

  • Biochemistry
  • Molecular Biology

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    Bannantine, J. P., Waters, W. R., Stabel, J. R., Palmer, M. V., Li, L., Kapur, V., & Paustian, M. L. (2008). Development and use of a partial Mycobacterium avium subspecies paratuberculosis protein array. Proteomics, 8(3), 463-474. https://doi.org/10.1002/pmic.200700644