Pseudomonas streptomycin resistance transposon associated with R-plasmid mobilization

Abstract

Plasmid pMG1 encodes resistance to gentamicin, streptomycin, sulfonamides, and mercuric ions and also mobilizes pRO161, a transfer-deficient plasmid derived from RP1. Upon mobilization, pRO161 acquires streptomycin resistance (Sm(r)) and can subsequently be remobilized by pMG1 at significantly higher frequencies than pRO161 itself. Both the initial acquisition of Sm(r) and the subsequent mobilization of the transfer-deficient plasmid are recA independent: thus, the Sm(r) determinant appears to be located on a transposon, designated Tn904. Tn904 transposes to a variety of other plasmids, including RP1, FP2, R388, K, pRO1600, and pBR322, and in some cases the acquisition of this transposon accompanied deletions in the target plasmid. When no deletion occurred, target plasmids gained 5.2 kilobase pairs of DNA and new restriction endonuclease cleavage sites for AvaI, BglII, PstI, SmaI, and SstI. Physical analysis of such plasmids showed that the Tn904 termini are inverted repeat DNA sequences of approximately 124 base pairs. After cloning into vector pRO1723, a single site for restriction endonuclease AvaI was identified within the Sm(r) determinant of Tn904. In Escherichia coli, but not in Pseudomonas aeruginosa, Tn904 shows a gene dosage-dependent expression of streptomycin resistance.