The recovery of Mycobacterium avium subspecies paratuberculosis from the intestine of infected ruminants for proteomic evaluation.
Johne's disease is a slowly developing intestinal disease, primarily of ruminants, caused by Mycobacterium avium subspecies paratuberculosis. The disease contributes to significant economic losses worldwide in agricultural industry. Analysis of bacterial proteomes isolated directly from infected animals can provide important information about the repertoire of proteins present during infection and disease progression. In this study, M. avium subspecies paratuberculosis has been extracted from Johne's disease-infected cattle and goat intestinal tissue sections in a manner compatible with direct 2-DE proteomic analysis for comparison with in vitro-cultured bacteria. M. avium subspecies paratuberculosis was harvested from the submucosa and mucosa of intestinal sections and enriched from macerated tissue by hypotonic lysis, sonication and centrifugation through a viscosity gradient. Subsequent comparison of the proteomes of the in vivo- and in vitro-derived bacteria identified a number of proteins that were differentially expressed. Among them, a number of hypothetical proteins of unknown function and a hypothetical fatty acyl dehydrogenase (FadE3_2) and 3-hydroxyacyl-CoA dehydrogenase, possibly important for in vivo metabolism, utilising the pathway for the beta-oxidation of fatty acids.
Protein Biochemistry and Proteomics Group, Australian Animal Health Laboratory, CSIRO Livestock Industries, Geelong VIC 3220, Australia.
Sharon
S
Egan
Mark
M
Lanigan
Brian
B
Shiell
Gary
G
Beddome
David
D
Stewart
Jill
J
Vaughan
Wojtek P
WP
Michalski
D000818:Animals; D001426:Bacterial Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D002417:Cattle; D002418:Cattle Diseases / Q000382:microbiology*; D015180:Electrophoresis, Gel, Two-Dimensional; D007422:Intestines / Q000382:microbiology*; D008969:Molecular Sequence Data; D016927:Mycobacterium avium subsp. paratuberculosis / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification* / Q000378:metabolism; D010283:Paratuberculosis / Q000382:microbiology*; D040901:Proteomics
D001426:Bacterial Proteins