Characterization of a tobacco Bright Yellow 2 cell line expressing the tetracycline repressor at a high level for strict regulation of transgene expression.
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.
David
K M
KM
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.
Perrot-Rechenmann
C
C
D016960:Agrobacterium tumefaciens / Q000235:genetics; D002460:Cell Line; D005966:Glucuronidase / Q000032:analysis / Q000235:genetics*; D007700:Kinetics; D010947:Plants, Toxic; D011994:Recombinant Proteins / Q000032:analysis / Q000096:biosynthesis; D012097:Repressor Proteins / Q000235:genetics*; D013752:Tetracycline / Q000494:pharmacology; D014026:Nicotiana / Q000166:cytology / Q000235:genetics*; D014162:Transfection
D011994:Recombinant Proteins; D012097:Repressor Proteins; C034811:tetracycline resistance-encoding transposon repressor protein; D005966:Glucuronidase; D013752:Tetracycline