Open AccessCharacterization of a Tobacco Bright Yellow 2 Cell Line Expressing the Tetracycline Repressor at a High Level for Strict Regulation of Transgene Expression
Author(s) -
Karine David,
Catherine PerrotRechenmann
Publication year - 2001
Publication title -
plant physiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 3.554
H-Index - 312
eISSN - 1532-2548
pISSN - 0032-0889
DOI - 10.1104/pp.125.4.1548
Subject(s) - tetr , nicotiana tabacum , transgene , repressor , biology , cell culture , tetracycline , reporter gene , microbiology and biotechnology , gene expression , green fluorescent protein , gene , genetics , antibiotics
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.