Physical and Functional Characteristics of R-Factor Deoxyribonucleic Acid Segregated into Escherichia coli Minicells

Both fi(+) and fi(-) type R factors function as intact replicative units when segregated into Escherichia coli minicells. Hybridization studies demonstrate that at least 95% of the deoxyribonucleic acid (DNA) in R(+) minicells is episomal in origin. About half of the DNA can be extracted in a closed circular form and about 75% of the DNA is membrane associated. DNA, ribonucleic acid, and protein synthesis proceeds in R(+) minicells in contrast to R(-) minicell controls. The system offers a unique opportunity to study a relatively small replicative unit in a native cell environment and a simple means of isolating large quantities of episomal DNA.