Transposon display: a versatile method for transposon tagging

Methods Mol Biol. 2013:1057:239-50. doi: 10.1007/978-1-62703-568-2_17.

Abstract

Transposon tagging has been used successfully in a range of organisms for the cloning of mutants of interest. In species containing high copy numbers of transposable elements combined with a high transposition rate, forward cloning can be quite challenging and requires specific high-resolution methods. Here we detail an updated version of the Transposon Display technique, which allows visualization of large numbers of transposon-flanking sequences simultaneously in a highly robust and reproducible manner. This strategy was developed for the analysis of the transpositional behavior of the dTph1 transposon and for the forward cloning of mutants, particularly in the Petunia W138 background, individuals of which can contain >200 copies of the endogenous dTph1 element. The method is derived from the AFLP technique and can in principle easily be adapted to any system.

MeSH terms

  • DNA Primers / genetics
  • DNA Transposable Elements / genetics*
  • DNA, Plant / genetics
  • DNA, Plant / isolation & purification
  • DNA, Plant / metabolism
  • Genetic Engineering / methods*
  • Petunia / genetics
  • Polymerase Chain Reaction

Substances

  • DNA Primers
  • DNA Transposable Elements
  • DNA, Plant