TAILIEUCHUNG - Báo cáo khoa học: Improvement of a monopartite ecdysone receptor gene switch and demonstration of its utility in regulation of transgene expression in plants

In plants, regulation of transgene expression is typically accomplished through the use of inducible promoter systems. The ecdysone receptor (EcR) gene switch is one of the best inducible systems available to regulate transgene expression in plants. | ỊFEBS Journal Improvement of a monopartite ecdysone receptor gene switch and demonstration of its utility in regulation of transgene expression in plants Venkata S. Tavva1 2 Subba R. Palli1 Randy D. Dinkins3 and Glenn B. Collins2 1 Department of Entomology University of Kentucky Lexington KY USA 2 Plant and SoilSciences Department University of Kentucky Lexington KY USA 3 USDA-ARS Forage-AnimalProduction Research Unit Lexington KY USA Keywords ecdysone receptor gene regulation methoxyfenozide transgenic plants zinc finger protein Correspondence S. R. Palli Department of Entomology 1100 Nicholasville Road University of Kentucky Lexington KY 40546-0091 USA Fax 1 859 323 1120 Tel 1 859 257 4962 E-mail rpalli@ Received 28 December 2007 revised 26 February 2008 accepted 3 March 2008 doi In plants regulation of transgene expression is typically accomplished through the use of inducible promoter systems. The ecdysone receptor EcR gene switch is one of the best inducible systems available to regulate transgene expression in plants. However the monopartite EcR gene switches developed to date require micromolar concentrations of ligand for activation. We tested several EcR mutants that were generated by changing one or two amino acid residues in the highly flexible ligand-binding domain of Choristoneura fumiferana EcR CfEcR . Based on the transient expression assays we selected a double mutant V395I Y415E VY of CfEcR CfEcRVY for further testing in stable transformation experiments. The CfEcRVY mutant only slightly improved the induction characteristics of the two-hybrid gene switch whereas the CfEcRVY mutant significantly improved the induction characteristics of the monopartite gene switch VGCfEVY . The ligand sensitivity of the VGCfEVY switch was improved by 125-15 625-fold in different transgenic lines analyzed compared to the VGCfEWt switch. The utility of the VGCfEVY switch was tested by regulating the expression of an Arabidopsis

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