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Generation of single-copy T-DNA transformants in Arabidopsis by the CRE/loxP recombination-mediated resolution system

Sylvie De Buck UGent, Ingrid Peck UGent, Chris De Wilde UGent, Gordana Marjanac UGent, Jonah Nolf UGent, Annelies De Paepe UGent and Anna Depicker UGent (2007) PLANT PHYSIOLOGY. 145(4). p.1171-1182
abstract
We investigated whether complex T-DNA loci, often resulting in low transgene expression, can be resolved efficiently into single copies by CRE/loxP-mediated recombination. An SB-loxP T-DNA, containing two invertedly oriented loxP sequences located inside and immediately adjacent to the T-DNA border ends, was constructed. Regardless of the orientation and number of SB-loxP-derived T-DNAs integrated at one locus, recombination between the outermost loxP sequences in direct orientation should resolve multiple copies into a single T-DNA copy. Seven transformants with a complex SB-loxP locus were crossed with a CRE-expressing plant. In three hybrids, the complex T-DNA locus was reduced efficiently to a single-copy locus. Upon segregation of the CRE recombinase gene, only the simplified T-DNA locus was found in the progeny, demonstrating DNA had been excised efficiently in the progenitor cells of the gametes. In the two transformants with an inverted T-DNA repeat, the T-DNA resolution was accompanied by at least a 10-fold enhanced transgene expression. Therefore, the resolution of complex loci to a single-copy T-DNA insert by the CRE/loxP recombination system can become a valuable method for the production of elite transgenic Arabidopsis thaliana plants that are less prone to gene silencing.
Please use this url to cite or link to this publication:
author
organization
year
type
journalArticle (original)
publication status
published
subject
keyword
LOX-SITES, TOBACCO PLANTS, INTEGRATION PATTERNS, THALIANA ROOT EXPLANTS, SITE-SPECIFIC RECOMBINATION, GLUCURONIDASE ACCUMULATION LEVELS, SELECTABLE MARKER GENE, TRANSGENE EXPRESSION, CRE RECOMBINASE, PLANT TRANSFORMATION
journal title
PLANT PHYSIOLOGY
Plant Physiol.
volume
145
issue
4
pages
1171 - 1182
Web of Science type
Article
Web of Science id
000251396300010
JCR category
PLANT SCIENCES
JCR impact factor
6.367 (2007)
JCR rank
7/149 (2007)
JCR quartile
1 (2007)
ISSN
0032-0889
DOI
10.1104/pp.107.104067
language
English
UGent publication?
yes
classification
A1
copyright statement
I have transferred the copyright for this publication to the publisher
id
409438
handle
http://hdl.handle.net/1854/LU-409438
date created
2008-05-15 18:26:00
date last changed
2013-10-18 15:46:55
@article{409438,
  abstract     = {We investigated whether complex T-DNA loci, often resulting in low transgene expression, can be resolved efficiently into single copies by CRE/loxP-mediated recombination. An SB-loxP T-DNA, containing two invertedly oriented loxP sequences located inside and immediately adjacent to the T-DNA border ends, was constructed. Regardless of the orientation and number of SB-loxP-derived T-DNAs integrated at one locus, recombination between the outermost loxP sequences in direct orientation should resolve multiple copies into a single T-DNA copy. Seven transformants with a complex SB-loxP locus were crossed with a CRE-expressing plant. In three hybrids, the complex T-DNA locus was reduced efficiently to a single-copy locus. Upon segregation of the CRE recombinase gene, only the simplified T-DNA locus was found in the progeny, demonstrating DNA had been excised efficiently in the progenitor cells of the gametes. In the two transformants with an inverted T-DNA repeat, the T-DNA resolution was accompanied by at least a 10-fold enhanced transgene expression. Therefore, the resolution of complex loci to a single-copy T-DNA insert by the CRE/loxP recombination system can become a valuable method for the production of elite transgenic Arabidopsis thaliana plants that are less prone to gene silencing.},
  author       = {De Buck, Sylvie and Peck, Ingrid and De Wilde, Chris and Marjanac, Gordana and Nolf, Jonah and De Paepe, Annelies and Depicker, Anna},
  issn         = {0032-0889},
  journal      = {PLANT PHYSIOLOGY},
  keyword      = {LOX-SITES,TOBACCO PLANTS,INTEGRATION PATTERNS,THALIANA ROOT EXPLANTS,SITE-SPECIFIC RECOMBINATION,GLUCURONIDASE ACCUMULATION LEVELS,SELECTABLE MARKER GENE,TRANSGENE EXPRESSION,CRE RECOMBINASE,PLANT TRANSFORMATION},
  language     = {eng},
  number       = {4},
  pages        = {1171--1182},
  title        = {Generation of single-copy T-DNA transformants in Arabidopsis by the CRE/loxP recombination-mediated resolution system},
  url          = {http://dx.doi.org/10.1104/pp.107.104067},
  volume       = {145},
  year         = {2007},
}

Chicago
De Buck, Sylvie, Ingrid Peck, Chris De Wilde, Gordana Marjanac, Jonah Nolf, Annelies De Paepe, and Anna Depicker. 2007. “Generation of Single-copy T-DNA Transformants in Arabidopsis by the CRE/loxP Recombination-mediated Resolution System.” Plant Physiology 145 (4): 1171–1182.
APA
De Buck, S., Peck, I., De Wilde, C., Marjanac, G., Nolf, J., De Paepe, A., & Depicker, A. (2007). Generation of single-copy T-DNA transformants in Arabidopsis by the CRE/loxP recombination-mediated resolution system. PLANT PHYSIOLOGY, 145(4), 1171–1182.
Vancouver
1.
De Buck S, Peck I, De Wilde C, Marjanac G, Nolf J, De Paepe A, et al. Generation of single-copy T-DNA transformants in Arabidopsis by the CRE/loxP recombination-mediated resolution system. PLANT PHYSIOLOGY. 2007;145(4):1171–82.
MLA
De Buck, Sylvie, Ingrid Peck, Chris De Wilde, et al. “Generation of Single-copy T-DNA Transformants in Arabidopsis by the CRE/loxP Recombination-mediated Resolution System.” PLANT PHYSIOLOGY 145.4 (2007): 1171–1182. Print.