Plasmids from freshwater environments capable of IncQ retrotransfer are diverse and include pQKH54, a new IncP-1 subgroup archetype

Microbiology (Reading). 2006 Sep;152(Pt 9):2689-2701. doi: 10.1099/mic.0.28941-0.

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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

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

Substances

  • DNA Primers
  • DNA Probes
  • DNA, Bacterial
  • Retroelements
  • DNA Restriction Enzymes
  • Mercury