Use of a known plasmid and transposon as tracers for the incompatibility classification of a cryptic plasmid

We have developed a novel genetic technique by which an unknown plasmid can be classified by the use of plasmid of known incompatibility group. In phenocopy state, cells harboring plasmids of known incompatibility group behave as normal recipients and receive plasmids belonging to the same incompatibility group at a high frequency. This work provides additional evidence that plasmids in 'phenocopy' hosts do not replicate and therefore fail to demonstrate their incompatibility barrier to the incoming plasmids. A cryptic plasmid, now called pWS7, has been identified by this method in a pili, fla female strain of E. coli K12. Genetic analysis shows the plasmid pWS7 is in fact, a sex-factor which is curable with acridine orange. It belongs to the Inc F1 group. Physical analysis confirms its size to be 124 Kb. The plasmid has been labelled genetically with a transposon Tn903 in a recA host and further characterized by heteroduplex analysis. A DNA sequence homology between pWS7 and F'lac plasmid extends only in F-regions, 2.8F-94.5F. The pili, fla host strain of pWS7 shows a high frequency of transformation for recombinant DNA and rapid propagation for a male-specific RNA phage, R17.