A new propionate (prp) locus of S. typhimurium was defined by mutation, was located to minute 8 of the chromosome, and was shown to be transcribed in the clockwise direction. A plasmid carrying the wild-type prp+ locus was isolated by complementation and its initial physical characterization is presented. Transcriptional regulation of prp was studied using MudI1734(lacZ+) operon fusions. Propionate stimulated prp transcription in a merodiploid strain containing prp+ and a prp::MudI1734 fusion, but failed to stimulate transcription of the same fusion in a haploid genetic background. prp transcription was reduced by a factor of 2 in strains deficient in the synthesis of the global regulatory protein FruR; fruR mutants failed to grow on propionate. Propionate blocked growth of prp mutants on medium containing succinate as carbon/energy source.