Plant Physiol, April 2001, Vol. 125, pp. 1548-1553
Characterization of a Tobacco Bright Yellow 2 Cell Line
Expressing the Tetracycline Repressor at a High Level for Strict
Regulation of Transgene Expression1
Karine M.
David and
Catherine
Perrot-Rechenmann*
Institut des Sciences Végétales, Centre National de la
Recherche Scientifique, Unité Propre de Recherche 040, Auxin
Perception and Transport Laboratory, Avenue de la Terrasse, 91198 Gif-sur-Yvette cedex, France
Manipulating the expression of a transgene in transient and stable
transformed cells is a requirement for many functional analyses. We
have investigated the use of the tetracycline-dependent gene expression
system developed by Gatz et al. (1992) in tobacco (Nicotiana
tabacum L. cv Bright Yellow 2 [BY2]) cells, the most widely
used plant cell culture. We have selected a BY2 cell line, named
BY2-tetracycline repressor (tetR) 17, which expresses the tetR at a
high level, and have evaluated the capacity of this cell line to
suppress the expression of a green fluorescent protein reporter gene
under the control of the "Triple-Op" promoter in the absence of
tetracycline in a large number of independent transformants. The
ability to induce the expression of green fluorescent protein after
treatment by anhydrotetracycline in the same transformants was also
analyzed. BY2-tetR17 cells were demonstrated to be excellent recipient
cells for recovery of clonal cell lines with a highly controlled
regulation of the introduced transgene.
1
K.D. was a recipient of the
Ministère de l'Education National, de la Recherche, et
de la Technologie Research Fellowship.
*
Corresponding author; e-mail rechenmann{at}isv.cnrs-gif.fr; fax
33-1-69-82-35-84.
© 2001 American Society of Plant Physiologists
