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Molecular Structure and Regulatory Potential of a T-DNA Integration Site in Petunia
Authors:Dietz-Pfeilstetter  Antje  Arndt  Nicola  Kay  Volker  Bode  Jürgen
Institution:(1) Federal Biological Research Centre for Agriculture and Forestry, Institute for Plant Virology, Microbiology and Biosafety, Messeweg 11/12, D-38104 Braunschweig, Germany;(2) German Research Center for Biotechnology (GBF), Epigenetic Regulation, Mascheroder Weg 1, D-38124 Braunschweig, Germany
Abstract:The genomic structure surrounding a T-DNA integration site in a transgenic petunia plant, which shows deregulation of a root-specific promoter, was investigated. We have already demonstrated that T-DNA integration in this transformant (P13) had occurred close to a scaffold/matrix attachment region (S/MAR). A major question regarding the observed promoter leakiness was whether the T-DNA had integrated into the centre or at the border of the Petun-SAR and whether other regulatory elements are located within this genomic region. While small rearrangements were shown to occur during T-DNA integration in agreement with other reports, we find indications of the presence of a SINE retroposon – an apparent landmark for recombinogenic targets – at the integration site. Binding assays to both plant and animal nuclear scaffolds, supported by biomathematical analyses, reveal that the T-DNA is definitely located at the border of a strong S/MAR, which is in agreement with current models on the structure of integration sites. These results, together with a developmentally regulated leaf-specific enhancer effect of the Petun-SAR on gene expression in transgenic tobacco plants, indicate that the Petun-SAR demarcates the right border of a chromatin domain with genes predominantly active in leaves.
Keywords:gene expression  Petunia hybrida  S/MAR  T-DNA integration  transgenic plants
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