Replicating function of the RS1 element associated with Vibrio cholerae CTXΦ prophage

The RS1 element associated with Vibrio choiei ne CTX Phi prophage was cloned from an El Tor biotype Vibrio cholerae strain. We used the recA(-) vaccine strain Peru-15, that lacks the target for RS-mediated site-specific integration. to show that RS1 promotes autonomous replication of a suicide vector. A linker insertion in the rstR open reading frame abolished autonomous replication in Peru-15 but not in a strain containing an RS1 in the chromosome. An AT-rich region containing cis-acting elements involved in autonomous replication was identified by deletion. This region was sufficient to support autonomous replication in a strain containing an RS1 in the chromosome. DNA sequence analysis of a region resent in RS1 and not RS2. revealed the presence of putative binding sites for host proteins involved in plasmid replication. These results indicate that RS1 contains a replicon distinct from RS2 which could be involved in replicative recombination events associated with tandem amplification of the CTX element. (C) 1998 Federation of European Microbiological Societies. Published by Elsevier Science B.V. All rights reserved.