Summary: Tn5Map, a Tn5 derivative containing the 18 bp I-cel site, was delivered from a RP4-mobilizable, RK6-derived suicide vector to HB101, and C58, which all lack natural I-Scel sites in their genomes. Digestion of the DNA from Tn5Map-containing strains and analysis by pulsed-field gel electrophoresis (PFGE) revealed that these derivatives contained a single transposon insertion. These digests also gave direct and independent proof for the single circular chromosome of , and for the presence of two circular chromosomes in and of a circular and a linear chromosome in C58 (which also contains two large circular plasmids). This rapid and versatile technique is potentially applicable to the study of the genomic organization in all Gram-negative bacteria which support Tn5 transposition. Moreover, linearization of circular replicons could be the first step for a rapid method of physical mapping.


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