Restriction enzyme fingerprinting of enterobacterial plasmids: a simple strategy with wide application

Abstract

SummaryRestriction enzyme fingerprints were generated from purified plasmid DNA from 324 clinical isolates that belonged to 7 enterobacterial genera and 88 single plasmids in Escherichia coli K12 according to the following strategy.Purified plasmid DNA was digested with PstI. The number of fragments detected in a 0·8 agarose gel was used to determine which 2 of 6 restriction enzymes including Pstl was most likely to provide a fingerprint comprising sufficient fragments to ensure specificity but sufficiently few to allow easy visual assessment and minimize coincidental matching. When PstI produced > 20 fragments, Eco RI and HindIII were used; when PstI generated < 6 fragments Bsp 1286 and AvaII were used and SmaI was employed when between 6 and 20 fragments were obtained from PstI digests. Using a minimum of 12 fragments from a combination of 2 enzymes as the criterion for characterizing a strain/plasmid, satisfactory 2-enzyme fingerprints were obtained from 87% of the strains and plasmids studied using PstI and no more than two additional enzymes per strain. Of the remaining 54 strains, 51 harboured only small plasmids (< 10 kb) and 3 produced satisfactory fingerprints when digested with a fourth enzyme.