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Lactic acid bacteria are frequently used in the preparation of fermented foods and as probiotics. They also are employed by the biotechnology industry to make valuable biomaterials, such as enzymes and lactic acid. The goal of this project was to produce shuttle vectors from the native plasmids found in different strains of Lactobacillus casei. Genetic regions from three native plasmids that contained the plasmid replication origin and genes encoding proteins involved with replication initiation and with partitioning of the plasmids into daughter cells were cloned into an Escherichia coli vector backbone. Maintenance of the resulting shuttle vectors in L. casei strain 12A was followed over 200 generations of growth in the absence of selection. The pDW8 vector, which was derived from a plasmid found in L. casei strain 12A, showed the best retention in these experiments. These vectors may prove useful for expression of foreign proteins in L. casei.

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