INVIVO DEFINITION OF THE FUNCTIONAL ORIGIN OF REPLICATION (ORI(+)) OF THE PROMISCUOUS PLASMID PLS1

We have defined the minimal origin of replication of the plasmid pLS1 leading strand, as comprised within a 247 bp region, by in vivo deletion analyses. Cloning of pLS1 DNA regions containing its oriV(+) into a compatible replicon resulted in weak incompatibility towards pLS1, but only when the cloned fragment included the entire pLS1 oriV(+). Plasmids lacking a functional repB gene (which encodes the pLS1 initiator of replication RepB protein) could be established in Streptococcus pneumoniae only when RepB was supplied in trans. We conclude that all the pLS1-encoded gene products involved in its replication and control are efficient trans-complementing plasmid elements.