The genetic and physical basis of variability in Escherichia coli strains carrying a reference Inc N group plasmid

The nature of basis of variability in the conjugational behaviour of RM98+ (RM98-carrying) strains of Escherichia coli K-12 that are otherwise similar in phenotype was studied. An explanation for such variability is provided. Some RM98+ strains of E. coli have a plasmid aggregate, which upon conjugation yields two different conjugative plasmids. The first (pCU1) is an N conjugative group plasmid by all available criteria. The second (pCU2) could not be placed in any conjugative group known among the Enterobacteriaceae. Reciprocal DNA hybridization experiments and the gel patterns displayed by the two plasmid DNAs upon digestion with different restriction endonucleases indicate no extensive sequence homology between pCU1 and pCU2. PCU2 DNA is much longer than pCU1 DNA. Despite the absence of extensive homology, the DNA of pCU1 and pCU2 can interact. Derivatives can be selected that have all the antibiotic markers of the aggregate plasmid but that neither contain nor segregate pCU2. It is shown that in such strains a DNA fragment of molecular weight 7.9 X 10(6) has been added to pCU1 concurrently with a tetracycline resistance marker originally present in pCU2 and absent in pCU1. These observations suggest that tetracycline resistance in pCU2 may be part of a large translocatable element. RM98 has been used to designate a reference Inc N group plasmid. The results presented indicate that this can lead to ambiguity. pCU1 would now be the appropriate reference plasmid.