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FLP/FRT Recombination from Yeast: Application of a Two Gene Cassette Scheme as an Inducible System in Plants
Department of Plant Sciences, University of Tennessee, Knoxville, TN 37996, USA
Biocentre Klein Flottbek, Developmental Biology and Biotechnology, University of Hamburg, Hamburg, Germany
* Author to whom correspondence should be addressed.
Received: 17 July 2010; in revised form: 23 August 2010 / Accepted: 6 September 2010 / Published: 13 September 2010
Abstract: Phytosensors are plants that are genetically engineered for sensing and reporting the presence of a specific contaminant, including agriculturally important biological agents. Phytosensors are constructed by transforming plants to contain specific biotic- or abiotic-inducible promoters fused to a reporter gene. When such transgenic plants encounter the target biotic or abiotic agent, the specific inducible promoter is triggered and subsequently drives the expression of the reporter gene, which produces a signal for detection. However, several systems lack robustness, rapid induction and promoter strength. Here, we tested the FLP/FRT recombination system in a construct containing a two gene cassette organization and examined its potential in transgenic Arabidopsis and tobacco plants using a b-glucuronidase (GUS) reporter. In this model system, a heat-shock inducible promoter was employed to control the expression of the FLP recombinase gene. Upon heat induction and subsequent active FLP-mediated excision event, the GUS gene was placed in close proximity to the 35S promoter resulting in an active GUS reporter expression. Our results demonstrate that the two gene cassette scheme of inducible FLP/FRT recombination system is functional in tobacco and Arabidopsis, providing additional insights into its possible application in phytosensing such as creating strong readout capabilities.
Keywords: FLP/FRT; site-specific recombination; heat shock; GUS reporter; tobacco; Arabidopsis; phytosensing
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MDPI and ACS Style
Rao, M.R.; Moon, H.S.; Schenk, T.M.H.; Becker, D.; Mazarei, M.; Stewart, C.N., Jr FLP/FRT Recombination from Yeast: Application of a Two Gene Cassette Scheme as an Inducible System in Plants. Sensors 2010, 10, 8526-8535.
Rao MR, Moon HS, Schenk TMH, Becker D, Mazarei M, Stewart CN, Jr. FLP/FRT Recombination from Yeast: Application of a Two Gene Cassette Scheme as an Inducible System in Plants. Sensors. 2010; 10(9):8526-8535.
Rao, Murali R.; Moon, Hong S.; Schenk, Tobias M. H.; Becker, Dirk; Mazarei, Mitra; Stewart, C. Neal, Jr. 2010. "FLP/FRT Recombination from Yeast: Application of a Two Gene Cassette Scheme as an Inducible System in Plants." Sensors 10, no. 9: 8526-8535.