Unique Salmonella choleraesuis surface protein affecting invasiveness: Possible inv related sequence

Abstract

TnphoA mutagenesis of a Salmonella choleraesuis isolate recovered from septicemic infection of feeder pigs resulted in 56 PhoA+ Kn(R) Str(R) mutants. Thirty-five mutants exhibited reduced levels of invasion in the Hep- 2 cell model and were examined by SDS-PAGE Western Blot analysis using an anti-alkaline phosphatase antibody to visualize the insertion gene products. A mutant which produced a gene fusion product of 95 kDa and exhibited >90% reduction in invasion was subcloned. A 10 Kb BamHI fragment of the chromosome containing the phoA insert was detected by hybridization and cloned into a pGEM vector. The resulting 1657 base sequence contained a 1104 bp ORF with two short regions of homology with S. typhimurium invF and invG, one region of homology with lcrD of Yersinia pseudotuberculosis but contained largely unique sequences not contained in Gene Bank. The full length sequence was not obtained as there was no stop codon detected. The % G+C was 44%, considerably lower than that of the Salmonella chromosome, but compatible with the proposed Yersinia origin of the inv genes. The NH2 387 a.a. sequence includes 5 transmembrane regions, resembling the model derived from the hydrophobicity plot of S. typhimurium InvA.