Double or triple sets of replication functions as inverted and direct repeats onin vitro reconstructed streptococcal MLS resistance plasmids

Abstract

In vitro rearrangement of plasmid pDB102 together with comparative studies of other streptococcal plasmids allowed the localization of replication and copy control functions on sequences which were present on pDB102 and its naturally occurring ancestor pSM19035 as duplicates in inverted orientation. Evidence is presented that neither the presence of duplicate replication regions nor their arrangement in inverted orientation was essential for plasmid survival. Among the in vitro reconstructed plasmids were several that stably carried two or three sets of replication and copy control functions either as inverted or direct repeats or both. A copy control mutation is described which led to a tenfold increase of copy number over that of the naturally occurring plasmid pSM19035.