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

Sylvie De Buck (UGent) , P Windels, Marc De Loose (UGent) and Anna Depicker (UGent)
(2004) CELLULAR AND MOLECULAR LIFE SCIENCES. 61(19-20). p.2632-2645
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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-beta-glucuronidase transgene expression, measured in two subsequent generations, was similar. This observation means that the intratransformant 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.
Keywords
T-DNA integration, position effect, transgene expression, AGROBACTERIUM-MEDIATED TRANSFORMATION, MATRIX ATTACHMENT REGIONS, VECTOR BACKBONE SEQUENCES, TRANSGENIC RICE PLANTS, EXPRESSION, THALIANA, METHYLATION, PATTERNS, PETUNIA, REPEATS

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Chicago
De Buck, Sylvie, P Windels, Marc De Loose, and Anna Depicker. 2004. “Single-copy T-DNAs Integrated at Different Positions in the Arabidopsis Genome Display Uniform and Comparable Β-glucuronidase Accumulation Levels.” Cellular and Molecular Life Sciences 61 (19-20): 2632–2645.
APA
De Buck, S., Windels, P., De Loose, M., & Depicker, A. (2004). Single-copy T-DNAs integrated at different positions in the Arabidopsis genome display uniform and comparable β-glucuronidase accumulation levels. CELLULAR AND MOLECULAR LIFE SCIENCES, 61(19-20), 2632–2645.
Vancouver
1.
De Buck S, Windels P, De Loose M, Depicker A. Single-copy T-DNAs integrated at different positions in the Arabidopsis genome display uniform and comparable β-glucuronidase accumulation levels. CELLULAR AND MOLECULAR LIFE SCIENCES. 2004;61(19-20):2632–45.
MLA
De Buck, Sylvie, P Windels, Marc De Loose, et al. “Single-copy T-DNAs Integrated at Different Positions in the Arabidopsis Genome Display Uniform and Comparable Β-glucuronidase Accumulation Levels.” CELLULAR AND MOLECULAR LIFE SCIENCES 61.19-20 (2004): 2632–2645. Print.
@article{299007,
  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-beta-glucuronidase transgene expression, measured in two subsequent generations, was similar. This observation means that the intratransformant 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.},
  author       = {De Buck, Sylvie and Windels, P and De Loose, Marc and Depicker, Anna},
  issn         = {1420-682X},
  journal      = {CELLULAR AND MOLECULAR LIFE SCIENCES},
  keywords     = {T-DNA integration,position effect,transgene expression,AGROBACTERIUM-MEDIATED TRANSFORMATION,MATRIX ATTACHMENT REGIONS,VECTOR BACKBONE SEQUENCES,TRANSGENIC RICE PLANTS,EXPRESSION,THALIANA,METHYLATION,PATTERNS,PETUNIA,REPEATS},
  language     = {eng},
  number       = {19-20},
  pages        = {2632--2645},
  title        = {Single-copy T-DNAs integrated at different positions in the Arabidopsis genome display uniform and comparable β-glucuronidase accumulation levels},
  url          = {http://dx.doi.org/10.1007/s00018-004-4284-8},
  volume       = {61},
  year         = {2004},
}

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