Nature Biotechnology
19, 157 - 161 (2001)
doi:10.1038/84428
Chemical-regulated, site-specific DNA excision in transgenic plantsJianru Zuo1, 2, Qi-Wen Niu1, 2, Simon Geir Møller1
& Nam-Hai Chua11
Laboratory of Plant Molecular Biology, The Rockefeller University, 1230 York Avenue, New York, NY 10021. 2
These authors contributed equally to this work.
Correspondence should be addressed to Nam-Hai Chua chua@rockvax.rockefeller.eduDNA excisionCre/loxPmarker-free transformantstransgenic plantsXVEWe have developed a chemical-inducible, site-specific DNA excision system in transgenic Arabidopsis plants mediated by the Cre/loxP DNA recombination system. Expression of the Cre recombinase was tightly controlled by an estrogen receptor-based fusion transactivator XVE. Upon induction by  -estradiol, sequences encoding the selectable marker, Cre, and XVE sandwiched by two loxP sites were excised from the Arabidopsis genome, leading to activation of the downstream GFP (green fluorescent protein) reporter gene. Genetic and molecular analyses indicated that the system is tightly controlled, showing high-efficiency inducible DNA excision in all 19 transgenic events tested with either single or multiple T-DNA insertions. The system provides a highly reliable method to generate marker-free transgenic plants after transformation through either organogenesis or somatic embryogenesis.
|
