Immunoblot reactivity of polyclonal and monoclonal antibodies with periplasmic flagellar proteins FlaA1 and FlaB of porcine Serpulina species.
The periplasmic-flagellum (PF) proteins of Triton X-100-soluble and Triton X-100-insoluble sodium dodecyl sulfate-treated fractions from reference and field strains of Serpulina hyodysenteriae, Serpulina innocens, and Serpulina pilosicoli were characterized by Western blotting with a rabbit polyclonal antibody (PAb) specific for the 44-kDa PF sheath protein of S. hyodysenteriae (Z. Li, F. Dumas, D. Dubreuil, and M. Jacques, J. Bacteriol. 175:8000-8007, 1993) and a murine monoclonal antibody (MAb), designated 7G2, specific for the PF core FlaB proteins of S. hyodysenteriae. The MAb 7G2 reacted with a conserved epitope present in the 37-, 34-, and 32-kDa PF core FlaB proteins of all Serpulina species. This suggested that the core FlaB proteins are conserved among porcine Serpulina species. An immunoreactive band of approximately 44 kDa was present with all S. hyodysenteriae, S. innocens, and S. pilosicoli strains that were reacted with the PAb. The specificities of the PAb and the MAb for the FlaA1 and FlaB proteins of Serpulina species were confirmed by N-terminal amino acid sequencing of 44- and 37-kDa proteins, respectively, of S. hyodysenteriae and S. pilosicoli. Results from this study provide further evidence that the 44-kDa protein FlaA1 and the 37-, 34-, and 32-kDa FlaB proteins are conserved among porcine Serpulina species.
Fisher
L N
LN
Department of Veterinary and Biomedical Sciences, University of Nebraska-Lincoln 68583-0905, USA.
Duhamel
G E
GE
Westerman
R B
RB
Mathiesen
M R
MR
D000595:Amino Acid Sequence; D000818:Animals; D000911:Antibodies, Monoclonal; D001426:Bacterial Proteins / Q000032:analysis / Q000276:immunology; D015153:Blotting, Western / Q000379:methods; D017400:Brachyspira / Q000302:isolation & purification* / Q000648:ultrastructure; D005407:Flagella / Q000737:chemistry / Q000276:immunology; D005408:Flagellin / Q000032:analysis / Q000737:chemistry / Q000276:immunology*; D051379:Mice; D013145:Spirochaetales Infections / Q000175:diagnosis*; D013552:Swine
D000911:Antibodies, Monoclonal; D001426:Bacterial Proteins; D005408:Flagellin; C067138:flaA protein, bacteria; C416532:flaB flagellin