Stringent control of transgene expression in Arabidopsis thaliana using the Top10 promoter system

Citation
J. Love et al., Stringent control of transgene expression in Arabidopsis thaliana using the Top10 promoter system, PLANT J, 21(6), 2000, pp. 579-588
Citations number
43
Categorie Soggetti
Plant Sciences","Animal & Plant Sciences
Journal title
PLANT JOURNAL
ISSN journal
09607412 → ACNP
Volume
21
Issue
6
Year of publication
2000
Pages
579 - 588
Database
ISI
SICI code
0960-7412(200003)21:6<579:SCOTEI>2.0.ZU;2-X
Abstract
We show that the tightly regulated tetracycline-sensitive Top10 promoter sy stem (Weinmann et al. Plant J. 1994, 5, 559-569) is functional in Arabidops is thaliana. A pure breeding A. thaliana line (JL-tTA/8) was generated whic h expressed a chimeric fusion of the tetracycline repressor and the activat ion domain of Herpes simplex virus (tTA), from a single transgenic locus. P lants from this line were crossed with transgenics carrying the ER-targeted green fluorescent protein coding sequence (mGFP(5)) under control of the T op10 promoter sequence. Progeny from this cross displayed ER-targeted GFP f luorescence throughout the plant, indicating that the tTA-Top10 promoter in teraction was functional in A. thaliana. GFP expression was repressed by 10 0 ng ml(-1) tetracycline, an order of magnitude lower than the concentratio n used previously to repress expression in Nicotiana tabacum. Moreover, the level of GFP expression was controlled by varying the concentration of tet racycline in the medium, allowing a titred regulation of transgenic activit y that was previously unavailable in A. thaliana. The kinetics of GFP activ ity were determined following de-repression of the Top10::mGFP(5) transgene , with a visible ER-targeted GFP signal appearing from 24 to 48 h after de- repression.