Indirect selection of bacterial plasmids lacking identifiable phenotypic properties

Abstract

A procedure is described that uses an indicator plasmid (pSC201) to identify cells in a bacterial population that have been co transformed with a second plasmid lacking detectable phenotypic properties. Under appropriate conditions of indirect selection, between 50 and 85% of transformants carrying the indicator plasmid also contain the nonselected plasmid. A temperature sensitive mutation in the replication functions of the indicator plasmid enables its elimination from doubly transformed bacteria. Using this procedure, the authors have isolated bacteria that carry only the small cryptic plasmid, P15A, of the Escherichia coli strain 15. This genetic element, which contains only 2,300 nucleotide pairs, is thus capable of functioning as a replicon independently of the two larger plasmids normally associated with it in E. coli 15 strains.