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Functional and comparative analysis of two distinct ecdysteroid-responsive gene expression constructs in Drosophila S2 cells.

Jeroen Poels·Alberto Martinez·Marie-Marthe Suner·Arnold De Loof·Stuart J Dunbar·Jozef Vanden Broeck
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Abstract

Inducible expression systems have proven to be of major interest when analysing the function of specific genes or when expressing cytotoxic proteins. In an effort to develop inducible switches allowing for flexible fine-tuning of gene expression levels in insect cells, we have compared the induction capacities of two Drosophila minimal promoters when linked to four consecutive ecdysone response elements. These minimal promoters, either containing a TATA-box or a downstream promoter element, drove the expression of a luciferase reporter gene. Potent induction capacities were observed with the insect moulting hormone, 20-hydroxyecdysone, and with ponasterone A, a plant ecdysteroid. The developed inducible switches further expand the repertoire of molecular tools for functional expression of proteins of interest in insect cells. In addition, the combination of an ecdysone switch with promoters that possess different structural elements can provide novel insights into ecdysteroid-induced transcription in an insect cell line.

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AnimalsBase SequenceCell LineDNA-Binding ProteinsDose-Response Relationship, DrugDrosophilaDrosophila ProteinsEcdysteroneGene Expression RegulationGenes, ReporterHSP70 Heat-Shock ProteinsInsect ProteinsLuciferasesMolecular Sequence DataPromoter Regions, GeneticReceptors, SteroidTranscription FactorsTransfectionComparative StudyJournal Article

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Jeroen Poels, Alberto Martinez, Marie-Marthe Suner, Arnold De Loof, Stuart J Dunbar, Jozef Vanden Broeck. 2004. Functional and comparative analysis of two distinct ecdysteroid-responsive gene expression constructs in Drosophila S2 cells.. https://doi.org/10.1016/j.ibmb.2004.02.005

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