• Cécile Collonnier
  • Alexandra Schattner
  • Georges Berthier
  • Francine Boyer
  • Géraldine Coué-Philippe
  • Annick Diolez
  • Marie-Noëlle Duplan
  • Sophie Fernandez
  • Naïma Kebdani
  • André Kobilinsky
  • Marcel Romaniuk
  • Marc de Beuckeleer
  • Marc de Loose
  • Pieter Windels
  • Yves Bertheau

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T25 is one of the 4 maize transformation events from which commercial lines have so far been authorized in Europe. It was created by polyethylene glycol-mediated transformation using a construct bearing one copy of the synthetic pat gene associated with both promoter and terminator of the 35S ribosomal gene from cauliflower mosaic virus. In this article, we report the sequencing of the whole T25 insert and the characterization of its integration site by using a genome walking strategy. Our results confirmed that one intact copy of the initial construct had been integrated in the plant genome. They also revealed, at the 5' junction of the insert, the presence of a second truncated 35S promoter, probably resulting from rearrangements which may have occurred before or during integration of the plasmid DNA. The analysis of the junction fragments showed that the integration site of the insert presented high homologies with the Huck retrotransposon family. By using one primer annealing in the maize genome and the other in the 5' end of the integrated DNA, we developed a reliable event-specific detection system for T25 maize. To provide means to comply with the European regulation, a real-time PCR test was designed for specific quantitation of T25 event by using Taqman chemistry.
Original languageEnglish
JournalJournal of AOAC International
Volume88
Issue number2
Pages (from-to)536-46
Number of pages11
ISSN1060-3271
Publication statusPublished - 2005

    Research areas

  • 3' Flanking Region, 5' Flanking Region, Base Sequence, DNA Primers, DNA, Plant, European Union, Food, Genetically Modified, Legislation, Food, Molecular Sequence Data, Promoter Regions, Genetic, Reproducibility of Results, Reverse Transcriptase Polymerase Chain Reaction, Seeds, Zea mays

ID: 552511