Selected translocation of plasmid genes: frequency and regional specificity of translocation of the Tn3 element

Abstract

A procedure is described that selects for the insertion of transposable antibiotic resistance elements in a variety of recipient replicons. The selected translocation procedure, which employs a plasmid having a temperature-sensitive defect in replication as a donor of transposable genetic elements, was used to investigate certain characteristics of the translocation process. Translocation of the Tn3 element from plasmid to plasmid apparently occurs at a 103 to 104-times-higher frequency than from plasmid to chromosome. In both cases, continued accumulation of Tn3 on recipient genomes is prevented by development of an apparent equilibrium when only a small fraction of molecules in the recipient population contain Tn3. An alternative method for estimation of translocation frequency showed that the translocation process is temperature sensitive and that its frequency is unaffected by the presence of host recA mutation. Insertions of Tn3 onto the 65 .times. 106-dalton R6-5 plasmid in Escherichia coli are clustered on EcoRI fragments 3 (8 of 23 insertions) and 9 (7 of 23 insertions), which contain 12 and 5%, respectively, of the R6-5 genome. The occurrence of multiple insertions of Tn3 within EcoRI fragment 9, which contains the IS[insertion sequence]1 element and a terminus of the Tn4 element, is consistent with earlier evidence indicating that terminal DNA sequences of already present transposable elements may provide recognition sequences for subsequent illegitimate recombinational events.