Relationship of Group P1 Plasmids Revealed by Heteroduplex Experiments: RP1, RP4, R68 and RK2 Are Identical

Abstract

The molecular relationships of the IncP1 plasmids, RP1, RP4, R68 and RK2 were tested by EM examination of heteroduplexes. In several hybridization experiments molecules were detected which had a 7.8% portion of incomplete reannealing. This heterologous region could be explained by the typical renaturation behavior of the transposon Tn1. The identity of the Tn1 transposon present in RP1 and RP4 was proved by heteroduplex experiments with .lambda. phage DNA containing this transposon. These results indicated that the plasmids RP1 and RP4 are identical. Additional heteroduplex experiments between plasmids R68.45 and RP8 and between R68.45 and RK2 were performed. R68.45, a derivative of R68, has a small DNA insertion and RP8 can be regarded as a large insertion mutant of RP4; both insertions were used as single-stranded hybridization markers. From the hybrid molecules formed, it was deduced that R68 and RK2 are identical with RP1 and RP4 as far as molecular structure is revealed by the technique used. [Escherichia coli and Pseudomonas aeruginosa were used.].