Construction of a Tn5 derivative determining resistance to gentamicin and spectinomycin using a fragment cloned from R1033.
A physical and genetic map of the IncP plasmid R1033 was constructed: restriction fragments were subcloned and antibiotic resistance genes were located. The map is consistent with previous reports that R1033 is a derivative of RP4 carrying a 16-kb transposon Tn1696 which contains the antibiotic-resistance determinants present on R1033 but not on RP4. A BamHI fragment from R1033, determining resistance to gentamicin, spectinomycin and streptomycin, was cloned into Tn5, replacing the central Bg/II fragment that determined kanamycin resistance, producing a recombinant transposon Tn5-GmSpSm. This was shown to transpose in Rhizobium leguminosarum at a frequency similar to that of the parental Tn5.
D004251:DNA Transposable Elements; D004269:DNA, Bacterial / Q000235:genetics; D004352:Drug Resistance, Microbial; D004926:Escherichia coli / Q000235:genetics*; D005798:Genes, Bacterial; D005822:Genetic Vectors; D005839:Gentamicins / Q000494:pharmacology*; D012231:Rhizobium / Q000235:genetics*; D000198:Spectinomycin / Q000494:pharmacology*; D014169:Transformation, Bacterial
D004251:DNA Transposable Elements; D004269:DNA, Bacterial; D005839:Gentamicins; D000198:Spectinomycin