Transposon-Aided Capture of Antibiotic Resistance Plasmids from Complex Samples

Methods Mol Biol. 2019:2016:151-157. doi: 10.1007/978-1-4939-9570-7_14.

Abstract

Transposon-aided capture of plasmids allows for antibiotic resistance plasmids to be acquired from complex samples. It is based on the insertion of a transposon, with a known origin of replication and selectable marker, into the plasmids present in a sample which can subsequently be captured after the removal of contaminating chromosomal DNA. Here we describe isolation of bacterial cells from a complex sample, DNA extraction, transposon-aided capture of plasmids in the sample, and analysis of the captured plasmids.

Keywords: Antibiotic resistance; Complex samples; EZ-Tn5; Plasmid; Transposon.

MeSH terms

  • Bacteria / genetics*
  • Bacteria / isolation & purification
  • Bacterial Infections / drug therapy
  • Bacterial Infections / microbiology
  • DNA Transposable Elements*
  • DNA, Bacterial / genetics
  • DNA, Bacterial / isolation & purification
  • Drug Resistance, Microbial*
  • Humans
  • Plasmids / genetics*
  • Plasmids / isolation & purification

Substances

  • DNA Transposable Elements
  • DNA, Bacterial