Viktorinová Ivana, Wimmer Ernst A
Department of Genetics, University of Bayreuth, Universitätsstrasse 30 NW I, 95447 Bayreuth, Germany.
Insect Biochem Mol Biol. 2007 Mar;37(3):246-54. doi: 10.1016/j.ibmb.2006.11.010. Epub 2006 Dec 5.
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.
二元表达系统对于通过过表达或错误表达进行功能基因分析至关重要。此类系统在多种生物体中的应用将有助于研究许多在模式生物中无法解决的生物学问题。在此，我们报告了一组构建体以及一种有效的动力学方法，用于定量比较基于GAL4/UAS、LexA/(LL)(4)和四环素控制的tTA/TRE的一系列不同二元表达系统。通过使用这些构建体，我们能够表明，在黑腹果蝇中，源自酵母的GAL4/UAS系统在激活应答基因表达方面比源自细菌的LexA/(LL)(4)和tTA/TRE系统更有效。这些构建体嵌入基于piggyBac的广泛转座子载体中，并且反式激活因子由广泛适用的3xP3启动子驱动。因此，这些构建体应该可以转移，以评估二元表达系统在非模式昆虫物种中的功能。
