1887

Abstract

Previously, a gene encoding a predicted 67 kDa membrane-associated ATPase subunit of the Clp protease (ClpX) was identified in a porcine strain (95/1000) of the intestinal spirochaete In the current study, the distribution of this large gene was investigated in a collection of strains representing all seven spp. Using PCR with internal primers, an 878 bp portion of the gene was detected in 29 of 35 strains (83 %) of , 6 of 24 strains (25 %) of , 14 of 16 strains (88 %) of , 6 of 17 strains (35 %) of , 1 of 6 strains (17 %) of , 1 of 2 strains (50 %) of and not in the single strain of . The whole gene was sequenced from 20 spp. strains and compared with the gene from 95/1000 (GenBank accession no. AY466377). The genes had 99.3–99.7 % nucleotide sequence similarity and the predicted products had 99.7–100 % amino acid sequence similarity. The gene from WesB, a human strain of , was cloned and expressed as a histidine-tagged fusion protein in BL21. The purified protein was used to vaccinate mice and their sera were found to recognize the expected ∼67 kDa protein in whole-cell preparations of WesB. Sera from mice vaccinated with formalin-treated whole-cell proteins of WesB reacted with the recombinant protein. These results indicate that ClpX is both conserved and immunogenic and hence might be useful as a subunit vaccine component for spp. infections. Sera from humans with no known exposure to reacted with the recombinant ClpX protein, indicating that it is unlikely to be useful as a reagent for serological detection of spp. infections.

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2007-07-01
2021-10-24
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