Production of pediocin in Pediococcus acidilactici is associated with pMBR 1.0, which encodes prepediocin, a pediocin immunity protein, and two proteins involved in secretion and precursor processing. These four genes are organized as an operon under control of a single promoter. We have constructed shuttle vectors that contain all four structural genes, the chromosomal promoter STP2201 from Streptococcus thermophilus, and repA from the 2-kbp S. thermophilus plasmid pER8. The recombinant plasmid, pPC318, expressed and secreted active pediocin in Escherichia coli. Streptococcus thermophilus, Lactococcus lactis subsp. lactis, and Enterococcus faecalis::were electrotransformed with pPC418, a modified vector fitted with an erythromycin resistance tracking gene. Pediocin was produced and secreted in each of the lactic acid bacteria, and production was Stable for up to ten passages. The expression of pediocin in dairy fermentation microbes has important implications for bacteriocins as food preservatives in dairy products.