Comparative analysis of binary expression systems for directed gene expression in transgenic insects |
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| Authors: | Viktorinová Ivana Wimmer Ernst A |
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| Affiliation: | 1. Rothamsted Research, Biological Chemistry and Crop Protection Department, Harpenden, UK;2. College of Life and Environmental Sciences, Biosciences, University of Exeter, Penryn Campus, Penryn, Cornwall TR10 9FE, UK;1. Programa de Pós-Graduação em Entomologia, Departamento de Entomologia, Universidade Federal de Viçosa, Brazil;2. Departamento de Biologia Geral, Universidade Federal de Viçosa, Brazil;3. Centro de Ciências Agrárias, Departamento de Biologia, Universidade Federal do Espírito Santo Alegre, ES, Brazil;1. Max-Planck-Institute of Neurobiology, 82152 Martinsried, Germany;2. Janelia Research Campus, Howard Hughes Medical Institute, Ashburn, VA 20147, USA |
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| Abstract: | Binary expression systems are of key interest to functional gene analysis by over- or misexpression. The application of such systems in diverse organisms would allow the study of many biological problems not addressable in model organisms. Here we report a set of constructs and an effective kinetic approach to quantitatively compare a series of diverse binary expression systems based on GAL4/UAS, LexA/(LL)(4) and tetracycline-controlled tTA/TRE. By the use of these constructs, we could show that in Drosophila melanogaster the yeast-derived GAL4/UAS systems are more effective in activating responder gene expression than the bacterial-derived LexA/(LL)(4) and tTA/TRE systems. The constructs are embedded in broad-range piggyBac-based transposon vectors and the transactivators are driven by the widely applicable 3xP3 promoter. These constructs should therefore be transferable to evaluate the functionality of binary expression systems in non-model insect species. |
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