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Microbiology (Reading) ; 152(Pt 9): 2689-2701, 2006 Sep.
Article in English | MEDLINE | ID: mdl-16946264

ABSTRACT

Nine mercury-resistance plasmids isolated from river epilithon were assessed for their ability to retrotransfer the non-conjugative IncQ plasmid, R300B, derivatives of which have commercial uses that may result in accidental or deliberate release into the environment. Retrotransfer frequencies ranging from 2.1 x 10(-4) to 1.75 x 10(-5) were obtained for five of the nine plasmids--the remaining plasmids showed low or undetectable retrotransfer ability. The majority of the retrotransfer-proficient plasmids could not be classified by the tests used. Classical incompatibility testing with RP4 identified pQKH6, pQKH54 and pQM719 as IncP-1. Hybridization to replicon probes confirmed this for pQKH6 and pQM719 and added pQKH33. PCR with primers designed to amplify trfA and korA regions of IncP-1 plasmids did not identify any other plasmids. Plasmids pQKH6 and pQM719 but not pQKH54 produced similar SphI restriction profiles to the IncP-1beta subgroup. The complete nucleotide sequence of pQKH54 was determined, revealing it to have a complete IncP-1 backbone but belonging to a new distinct subgroup which was designated IncP-1gamma. The results emphasize the ubiquity and diversity of IncP-1 plasmids in the environment but demonstrate that plasmids of as yet unknown groups are also able to retrotransfer IncQ plasmids efficiently.


Subject(s)
Conjugation, Genetic , Environment , Fresh Water , Plasmids/genetics , Plasmids/isolation & purification , Base Composition , Base Sequence , DNA Primers , DNA Probes , DNA Restriction Enzymes/metabolism , DNA, Bacterial/genetics , Drug Resistance, Bacterial/genetics , Mercury/pharmacology , Models, Genetic , Molecular Sequence Data , Nucleic Acid Amplification Techniques , Nucleic Acid Hybridization , Phylogeny , Plasmids/classification , Polymerase Chain Reaction , Replication Origin , Replicon , Restriction Mapping , Retroelements , Sequence Analysis, DNA
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