title: Sequence polymorphisms in a surface PPE protein distinguish types I, II, and III of Mycobacterium avium subsp. paratuberculosis.
abstract: In the last 2 decades, a variety of different molecular typing methods have been developed to differentiate strains of Mycobacterium avium subsp. paratuberculosis. The most successful techniques are based on insertion sequences, repetitive loci, comparative genomics, or single nucleotide polymorphisms. In the present study, we chose to examine whether a single M. avium subsp. paratuberculosis gene could serve as a means of differentiation of a variety of isolates. The MAP1506 gene locus encodes a member of the polymorphic PPE protein family that has putative roles relevant to M. avium subsp. paratuberculosis pathogenicity. The MAP1506 locus was sequenced from a collection of 58 M. avium subsp. paratuberculosis isolates from different sources, hosts, and typing profiles. Following sequence alignment and analysis, it was found that bovine (type II) strains of M. avium subsp. paratuberculosis consistently differed from ovine (type I) and intermediate (type III) strains in seven and eight nucleotides, respectively. Polymorphic regions of the MAP1506 locus were selected for analysis by denaturing gradient gel electrophoresis, allowing visual discrimination of the three subtypes of M. avium subsp. paratuberculosis isolates. This is the first report describing the use of PCR and denaturing gradient gel electrophoresis on a single gene as a method to distinguish types I, II, and III of M. avium subsp. paratuberculosis.
authors.affiliation: Department of Production Animal Health, Faculty of Veterinary Medicine, University of Calgary, 3330 Hospital Drive NW, Calgary, Alberta T2N 4N1, Canada.
authors.forename: Tanya A
authors.initials: TA
authors.lastname: Griffiths
authors.forename: Kevin
authors.initials: K
authors.lastname: Rioux
authors.forename: Jeroen
authors.initials: J
authors.lastname: De Buck
mesh_terms: D000818:Animals; D001426:Bacterial Proteins / Q000737:chemistry / Q000235:genetics; D015373:Bacterial Typing Techniques; D002417:Cattle; D004251:DNA Transposable Elements; D004591:Electrophoresis, Polyacrylamide Gel / Q000379:methods; D008565:Membrane Proteins / Q000737:chemistry / Q000235:genetics*; D008969:Molecular Sequence Data; D016927:Mycobacterium avium subsp. paratuberculosis / Q000145:classification* / Q000235:genetics*; D010283:Paratuberculosis / Q000382:microbiology*; D012150:Polymorphism, Restriction Fragment Length; D020641:Polymorphism, Single Nucleotide; D017422:Sequence Analysis, DNA; D013045:Species Specificity
chemical_list: D001426:Bacterial Proteins; D004251:DNA Transposable Elements; D008565:Membrane Proteins