Identification and expression of a copy number control gene in the IncFIII hemolytic plasmid pSU316

Abstract

A DNA fragment carrying both the IncFIII determinant and a copy number control gene of the hemolytic plasmid pSU316 has been cloned in pBR322. Deletion derivatives of the hybrid plasmid generated by Bal 31 digestion, which no longer exhibit the IncFIII phenotype, fall into two complementation groups when tested against a pSU316 miniplasmid derivative. Type 1 mutants exhibit the copy number control (Cop+) phenotype whereas type 2 mutants do not. Restriction analysis of type 1 and type 2 mutants allowed us to locate the cop gene of pSU316 in a 700-base-pair fragment adjacent to the IncFIII determinant. Plasmid expression in a minicell system suggests that the product of the cop gene of pSU316 could be a 13,000-dalton protein.