This article examines alterations in a broad-host-range plasmid (pQSR50) that were observed following transfer to indigenous marine bacteria by natural transformation. Plasmid DNA from the transformants had altered restriction profiles. However, with the exception of the EcoRI site from one transformant (BS10), fragments amplified by polymerase chain reaction (PCR) and encompassing the recognition sites were cleaved by the relevant endonucleases, providing the sites were present. Analysis with DpnI and MboI indicated differences in DNA methylation between pQSR50 and the transformants. The missing EcoRI site from BS10 and smaller EcoRI fragments observed in transformants indicated that rearrangements had also occurred. Evolution of novel plasmid molecules following gene transfer may be an important mechanism by which natural genetic diversity is generated.