Advanced search
1 file | 291.84 KB Add to list

Increased leakiness of the tetracycline-inducible Triple-Op promoter in dividing cells renders it unsuitable for high inducible levels of a dominant negative CDC2aAt gene

Lieven De Veylder (UGent) , Tom Beeckman (UGent) , Marc Van Montagu (UGent) and Dirk Inzé (UGent)
(2000) JOURNAL OF EXPERIMENTAL BOTANY. 51(351). p.1647-1653
Author
Organization
Abstract
A tetracycline-inducible promoter system was used to generate transgenic tobacco plants that confer inducible expression of the wild type or a dominant negative allele of the gene coding for the cyclin-dependent kinase (CDK) of Arabidopsis thaliana CDC2aAt. Although the total extractable CDK activity was doubled, the induced expression of the wild-type CDC2aAt did not correlate with any change of the cell cycle kinetics. An increase of CDK activity upon CDC2aAt expression was only seen in dividing cell populations, demonstrating that CDC2aAt expression itself is not sufficient to induce CDK activation. Induced expression of the dominant negative CDC2aAt.N146 correlated with a reduction of CDK activity to 66% of the level found in non-induced cells. This decrease was not sufficient to block cell division. The isolation of plants showing only low inducible levels of CDC2aAt.N146 suggests that a counterselection against strong inducible lines had occurred. Accordingly, Triple-Op promoter activity was found in dividing cells in the absence of tetracycline.
Keywords
cell cycle, Arabidopsis thaliana, chemical-induced expression, cyclin-dependent kinase, TRANSGENIC TOBACCO PLANTS, CYCLIN-DEPENDENT KINASES, CAMV 35S PROMOTER, EXPRESSION SYSTEM, ARABIDOPSIS-THALIANA, TET REPRESSOR, WHOLE PLANTS, INDUCTION, PHOSPHORYLATION, HOMOLOG

Downloads

  • (...).pdf
    • full text
    • |
    • UGent only
    • |
    • PDF
    • |
    • 291.84 KB

Citation

Please use this url to cite or link to this publication:

MLA
De Veylder, Lieven, Tom Beeckman, Marc Van Montagu, et al. “Increased Leakiness of the Tetracycline-inducible Triple-Op Promoter in Dividing Cells Renders It Unsuitable for High Inducible Levels of a Dominant Negative CDC2aAt Gene.” JOURNAL OF EXPERIMENTAL BOTANY 51.351 (2000): 1647–1653. Print.
APA
De Veylder, L., Beeckman, T., Van Montagu, M., & Inzé, D. (2000). Increased leakiness of the tetracycline-inducible Triple-Op promoter in dividing cells renders it unsuitable for high inducible levels of a dominant negative CDC2aAt gene. JOURNAL OF EXPERIMENTAL BOTANY, 51(351), 1647–1653.
Chicago author-date
De Veylder, Lieven, Tom Beeckman, Marc Van Montagu, and Dirk Inzé. 2000. “Increased Leakiness of the Tetracycline-inducible Triple-Op Promoter in Dividing Cells Renders It Unsuitable for High Inducible Levels of a Dominant Negative CDC2aAt Gene.” Journal of Experimental Botany 51 (351): 1647–1653.
Chicago author-date (all authors)
De Veylder, Lieven, Tom Beeckman, Marc Van Montagu, and Dirk Inzé. 2000. “Increased Leakiness of the Tetracycline-inducible Triple-Op Promoter in Dividing Cells Renders It Unsuitable for High Inducible Levels of a Dominant Negative CDC2aAt Gene.” Journal of Experimental Botany 51 (351): 1647–1653.
Vancouver
1.
De Veylder L, Beeckman T, Van Montagu M, Inzé D. Increased leakiness of the tetracycline-inducible Triple-Op promoter in dividing cells renders it unsuitable for high inducible levels of a dominant negative CDC2aAt gene. JOURNAL OF EXPERIMENTAL BOTANY. 2000;51(351):1647–53.
IEEE
[1]
L. De Veylder, T. Beeckman, M. Van Montagu, and D. Inzé, “Increased leakiness of the tetracycline-inducible Triple-Op promoter in dividing cells renders it unsuitable for high inducible levels of a dominant negative CDC2aAt gene,” JOURNAL OF EXPERIMENTAL BOTANY, vol. 51, no. 351, pp. 1647–1653, 2000.
@article{124905,
  abstract     = {A tetracycline-inducible promoter system was used to generate transgenic tobacco plants that confer inducible expression of the wild type or a dominant negative allele of the gene coding for the cyclin-dependent kinase (CDK) of Arabidopsis thaliana CDC2aAt. Although the total extractable CDK activity was doubled, the induced expression of the wild-type CDC2aAt did not correlate with any change of the cell cycle kinetics. An increase of CDK activity upon CDC2aAt expression was only seen in dividing cell populations, demonstrating that CDC2aAt expression itself is not sufficient to induce CDK activation. Induced expression of the dominant negative CDC2aAt.N146 correlated with a reduction of CDK activity to 66% of the level found in non-induced cells. This decrease was not sufficient to block cell division. The isolation of plants showing only low inducible levels of CDC2aAt.N146 suggests that a counterselection against strong inducible lines had occurred. Accordingly, Triple-Op promoter activity was found in dividing cells in the absence of tetracycline.},
  author       = {De Veylder, Lieven and Beeckman, Tom and Van Montagu, Marc and Inzé, Dirk},
  issn         = {0022-0957},
  journal      = {JOURNAL OF EXPERIMENTAL BOTANY},
  keywords     = {cell cycle,Arabidopsis thaliana,chemical-induced expression,cyclin-dependent kinase,TRANSGENIC TOBACCO PLANTS,CYCLIN-DEPENDENT KINASES,CAMV 35S PROMOTER,EXPRESSION SYSTEM,ARABIDOPSIS-THALIANA,TET REPRESSOR,WHOLE PLANTS,INDUCTION,PHOSPHORYLATION,HOMOLOG},
  language     = {eng},
  number       = {351},
  pages        = {1647--1653},
  title        = {Increased leakiness of the tetracycline-inducible Triple-Op promoter in dividing cells renders it unsuitable for high inducible levels of a dominant negative CDC2aAt gene},
  url          = {http://dx.doi.org/10.1093/jexbot/51.351.1647},
  volume       = {51},
  year         = {2000},
}

Altmetric
View in Altmetric
Web of Science
Times cited: