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Plant Physiology logoLink to Plant Physiology
. 2001 Apr;125(4):1548–1553. doi: 10.1104/pp.125.4.1548

Characterization of a Tobacco Bright Yellow 2 Cell Line Expressing the Tetracycline Repressor at a High Level for Strict Regulation of Transgene Expression

K M David, C Perrot-Rechenmann
PMCID: PMC1539379  PMID: 11299335

Abstract

Abstract

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.


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