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Development and application of novel constructs to score C:G-to-T:A transitions and homologous recombination in Arabidopsis

Gert Van der Auwera UGent, Joke Baute UGent, Melanie Bauwens UGent, Ingrid Peck UGent, Denis Piette, Michael Pycke, Pieter Asselman UGent and Anna Depicker UGent (2008) PLANT PHYSIOLOGY. 146(1). p.22-31
abstract
We report on the development of five missense mutants and one recombination substrate of the beta-glucuronidase (GUS)encoding gene of Escherichia coli and their use for detecting mutation and recombination events in transgenic Arabidopsis (Arabidopsis thaliana) plants by reactivation of GUS activity in clonal sectors. The missense mutants were designed to find C:G-to-T:A transitions in a symmetrical sequence context and are in that respect complementary to previously published GUS point mutants. Small peptide tags (hemagglutinin tag and Strep tag II) and green fluorescent protein were translationally fused to GUS, which offers possibilities to check for mutant GUS production levels. We show that spontaneous mutation and recombination events took place. Mutagenic treatment of the plants with ethyl methanesulfonate and ultraviolet-C increased the number of mutations, validating the use of these constructs to measure mutation and recombination frequencies in plants exposed to biotic or abiotic stress conditions, or in response to different genetic backgrounds. Plants were also subjected to heavy metals, methyl jasmonate, salicylic acid, and heat stress, for which no effect could be seen. Together with an ethyl methanesulfonate mutation induction level much higher than previously described, the need is illustrated for many available scoring systems in parallel. Because all GUS missense mutants were cloned in a bacterial expression vector, they can also be used to score mutation events in E. coli.
Please use this url to cite or link to this publication:
author
organization
year
type
journalArticle (original)
publication status
published
subject
keyword
ACCUMULATION, FREQUENCY, GENOME, TRANSGENIC PLANTS, HUMAN BETA-GLUCURONIDASE, TRANSFORMATION, EXPRESSION, MUTATIONS, REVEALS, VECTORS
journal title
PLANT PHYSIOLOGY
Plant Physiol.
volume
146
issue
1
pages
22 - 31
Web of Science type
Article
Web of Science id
000252093100003
JCR category
PLANT SCIENCES
JCR impact factor
6.11 (2008)
JCR rank
12/155 (2008)
JCR quartile
1 (2008)
ISSN
0032-0889
DOI
10.1104/pp.107.105213
language
English
UGent publication?
yes
classification
A1
copyright statement
I have transferred the copyright for this publication to the publisher
id
409429
handle
http://hdl.handle.net/1854/LU-409429
date created
2008-05-15 18:21:00
date last changed
2013-10-21 11:09:54
@article{409429,
  abstract     = {We report on the development of five missense mutants and one recombination substrate of the beta-glucuronidase (GUS)encoding gene of Escherichia coli and their use for detecting mutation and recombination events in transgenic Arabidopsis (Arabidopsis thaliana) plants by reactivation of GUS activity in clonal sectors. The missense mutants were designed to find C:G-to-T:A transitions in a symmetrical sequence context and are in that respect complementary to previously published GUS point mutants. Small peptide tags (hemagglutinin tag and Strep tag II) and green fluorescent protein were translationally fused to GUS, which offers possibilities to check for mutant GUS production levels. We show that spontaneous mutation and recombination events took place. Mutagenic treatment of the plants with ethyl methanesulfonate and ultraviolet-C increased the number of mutations, validating the use of these constructs to measure mutation and recombination frequencies in plants exposed to biotic or abiotic stress conditions, or in response to different genetic backgrounds. Plants were also subjected to heavy metals, methyl jasmonate, salicylic acid, and heat stress, for which no effect could be seen. Together with an ethyl methanesulfonate mutation induction level much higher than previously described, the need is illustrated for many available scoring systems in parallel. Because all GUS missense mutants were cloned in a bacterial expression vector, they can also be used to score mutation events in E. coli.},
  author       = {Van der Auwera, Gert and Baute, Joke and Bauwens, Melanie and Peck, Ingrid and Piette, Denis and Pycke, Michael and Asselman, Pieter and Depicker, Anna},
  issn         = {0032-0889},
  journal      = {PLANT PHYSIOLOGY},
  keyword      = {ACCUMULATION,FREQUENCY,GENOME,TRANSGENIC PLANTS,HUMAN BETA-GLUCURONIDASE,TRANSFORMATION,EXPRESSION,MUTATIONS,REVEALS,VECTORS},
  language     = {eng},
  number       = {1},
  pages        = {22--31},
  title        = {Development and application of novel constructs to score C:G-to-T:A transitions and homologous recombination in Arabidopsis},
  url          = {http://dx.doi.org/10.1104/pp.107.105213},
  volume       = {146},
  year         = {2008},
}

Chicago
Van der Auwera, Gert, Joke Baute, Melanie Bauwens, Ingrid Peck, Denis Piette, Michael Pycke, Pieter Asselman, and Anna Depicker. 2008. “Development and Application of Novel Constructs to Score C:G-to-T:A Transitions and Homologous Recombination in Arabidopsis.” Plant Physiology 146 (1): 22–31.
APA
Van der Auwera, G., Baute, J., Bauwens, M., Peck, I., Piette, D., Pycke, M., Asselman, P., et al. (2008). Development and application of novel constructs to score C:G-to-T:A transitions and homologous recombination in Arabidopsis. PLANT PHYSIOLOGY, 146(1), 22–31.
Vancouver
1.
Van der Auwera G, Baute J, Bauwens M, Peck I, Piette D, Pycke M, et al. Development and application of novel constructs to score C:G-to-T:A transitions and homologous recombination in Arabidopsis. PLANT PHYSIOLOGY. 2008;146(1):22–31.
MLA
Van der Auwera, Gert, Joke Baute, Melanie Bauwens, et al. “Development and Application of Novel Constructs to Score C:G-to-T:A Transitions and Homologous Recombination in Arabidopsis.” PLANT PHYSIOLOGY 146.1 (2008): 22–31. Print.