Cellular and Molecular Life Sciences CMLS

, Volume 61, Issue 19–20, pp 2632–2645

Single-copy T-DNAs integrated at different positions in the Arabidopsis genome display uniform and comparable β-glucuronidase accumulation levels

  • S. De Buck
  • P. Windels
  • M. De Loose
  • A. Depicker
Research Article

Abstract.

This study aimed at determining whether transgene expression variability is observed in single-copy T-DNA plants and whether it can be correlated with the T-DNA integration position. Among a population of 135 Arabidopsis thaliana transformants, selected on the basis of antibiotic resistance marker expression, 21 single-copy T-DNA transformants were identified and characterized. In 19 of these 21 lines, 35S-β-glucuronidase transgene expression, measured in two subsequent generations, was similar. This observation means that the intra-transformant variability was as high as the inter-transformant variability. Integration into an intergenic or genic region, into an exon or intron, in sense or antisense orientation, did not result in differential transgene expression. Remarkably, single-copy transformants were not always the highest expressers, implying that low transgene expression is not always induced by multicopy transformants. In only 2 of the 21 single-copy plants was the transgene expression more than 20-fold lower. However, characteristics of the insertion position in one of these lines did not differ significantly when compared to high-expressing lines. In the remaining line, methylation of the transgene was clearly demonstrated. In conclusion, screening for single-copy T-DNA transformants greatly enriches for stable and high transgene expression, because the integration position is not a major determinant of transgene expression variability in Arabidopsis.

Key words.

Position effect T-DNA integration transgene expression 

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Copyright information

© Birkhäuser Verlag, Basel 2004

Authors and Affiliations

  • S. De Buck
    • 1
  • P. Windels
    • 2
  • M. De Loose
    • 2
  • A. Depicker
    • 1
  1. 1.Department of Plant Systems Biology, Flanders Interuniversity Institute for Biotechnology (VIB)Ghent UniversityGentBelgium
  2. 2.Department of Plant Genetics and BreedingCenter of Agricultural Research-GentMelleBelgium

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