Abstract
We have identified two 19-kb conjugative transposons (Tn5381 and Tn5383) in separate strains of multiply resistant Enterococcus faecalis. These transposons confer resistance to tetracycline and minocycline via a tetM gene, are capable of both chromosomal and plasmid integration in a Rec- environment, and transfer between strains in the absence of detectable plasmid DNA at frequencies ranging from < 1 x 10(-9) to 2 x 10(-5) per donor CFU, depending on the donor strain and the growth conditions. Hybridization studies indicate that these transposons are closely related to Tn916. We have identified bands of ca. 19 kb on agarose gel separations of alkaline lysis preparations from E. faecalis strains containing chromosomal copies of Tn5381, which we have confirmed to be a circularized form of this transposon. This phenomenon has previously been observed only when Tn916 has been cloned in Escherichia coli. Overnight growth of donor strains in the presence of subinhibitory concentrations of tetracycline results in an approximately 10-fold increase in transfer frequency of Tn5381 into enterococcal recipients and an increase in the amount of the circular form of Tn5381 as detectable by hybridization. These results suggest that Tn5381 is a Tn916-related conjugative transposon for which the appearance of a circular form and the conjugative-transfer frequency are regulated by a mechanism(s) affected by the presence of tetracycline in the growth medium.
MeSH Terms
Base Sequence
Conjugation, Genetic
Crosses, Genetic
DNA Transposable Elements
DNA, Bacterial/genetics,isolation & purification
DNA, Circular/analysis,genetics
Deoxyribonuclease EcoRI/metabolism
Enterococcus faecalis/drug effects,genetics
Molecular Sequence Data
Oligodeoxyribonucleotides
Plasmids
Polymerase Chain Reaction
Restriction Mapping
Tetracycline/pharmacology
Chemicals
DNA Transposable Elements
DNA, Bacterial
DNA, Circular
Oligodeoxyribonucleotides
Deoxyribonuclease EcoRI
Tetracycline
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Rice L B
Division of Infectious Disease, Department of Veteran's Affairs Medical Center, Cleveland, Ohio.
Marshall S H
Carias L L
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