Characterization of pRS5: a theta-type plasmid found in a strain of Pediococcus pentosaceus isolated from wine that can be used to generate cloning vectors for lactic acid bacteria.

The nucleotide sequence of pRS5 (10153bp) is reported. Through sequence analysis, 9 open reading frames (ORFs) were identified and the following features observed: a region likely involved in replication whose structural features indicate that pRS5 belongs to the pUCL287 group of theta-type replicons, and hypothetical proteins putatively involved in plasmid copy number control, restriction-modification system, toxin-antitoxin system and a putative integrase. Shuttle vectors for Escherichia coli and lactic acid bacteria (LAB) as well as a small cloning vector for direct use in LAB were constructed using the replication region of pRS5. The ability of such vectors to accept and express other genes was assessed. All pRS5-derivatives were maintained at a high rate over 200 generations without selective pressure.