Construction of a mariner-based transposon for epitope-tagging and genomic targeting

Gene. 2002 Aug 21;296(1-2):179-85. doi: 10.1016/s0378-1119(02)00856-9.


Mobile genetic elements are often employed for constructing gene fusions or to perform mutagenesis. mariner transposons are well-suited to such applications because of their low site specificity, in vitro activity, and exceptionally broad host range. This report describes a mariner-based method for rapidly creating a large number of insertion mutants that can be converted to in-frame epitope fusions in a single step. First, a mariner-based vector is used to deliver a FLP recombinase substrate randomly into a target molecule. Expression of the FLP recombinase is then induced to catalyse the excision of sequences flanked by FLP recombinase target recognition sites, leaving behind a triple-FLAG epitope. The reversibility of the excision event provides opportunities for using genomic targeting methods easily to create transcriptional or translational fusions to genes of interest.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Cloning, Molecular
  • DNA Nucleotidyltransferases / genetics
  • DNA Nucleotidyltransferases / metabolism
  • DNA Transposable Elements / genetics*
  • DNA, Bacterial / genetics
  • DNA, Bacterial / metabolism
  • Epitopes / genetics
  • Escherichia coli / genetics
  • Gene Targeting / methods*
  • Lac Operon / genetics
  • Mutagenesis, Insertional
  • Oligopeptides
  • Peptides / genetics*
  • Plasmids / genetics
  • Recombination, Genetic / genetics


  • DNA Transposable Elements
  • DNA, Bacterial
  • Epitopes
  • Oligopeptides
  • Peptides
  • FLAG peptide
  • DNA Nucleotidyltransferases
  • FLP recombinase