Slade F A, Staveley B E
Department of Biology, Faculty of Science, Memorial University of Newfoundland, St. John's, Newfoundland and Labrador, Canada.
Genet Mol Res. 2015 Oct 19;14(4):12788-96. doi: 10.4238/2015.October.19.22.
The UAS-Gal4 ectopic expression system is a widely used and highly valued tool that allows specific gene expression in Drosophila melanogaster. Yeast transcription factor Gal4 can be directed using D. melanogaster transcriptional control elements, and is often assumed to have little effect on the organism. By evaluation of the consequences of maternal and paternal inheritance of a Gal4 transgene under the transcriptional regulation of armadillo control elements (arm-Gal4), we demonstrated that Gal4 expression could be detrimental to development and longevity. Male progeny expressing arm-Gal4 in the presence of UAS-lacZ transgene had reduced numbers and size of ommatidia, compared to flies expressing UAS-lacZ transgene under the control of other Gal4 transgenes. Aged at 25°C, the median life span of male flies with maternally inherited elav-Gal4 was 70 days, without a responding transgene or with UAS-lacZ. The median life span of maternally inherited arm-Gal4 male flies without a responding transgene was 48 days, and 40 days with the UAS-lacZ transgene. A partial rescue of this phenotype was observed with the expression of UAS-lacZ under paternal arm-Gal4 control, having an average median lifespan of 60 days. This data suggests that arm-Gal4 has detrimental effects on Drosophila development and lifespan that are directly dependent upon parental inheritance, and that the benign responder and reporter gene UAS-lacZ may influence D. melanogaster development. These findings should be taken into consideration during the design and execution of UAS-Gal4 expression experiments.
UAS-Gal4异位表达系统是一种广泛使用且备受重视的工具,可在黑腹果蝇中实现特定基因的表达。酵母转录因子Gal4可通过黑腹果蝇转录控制元件进行定向表达,并且通常认为对生物体影响较小。通过评估在犰狳控制元件(arm-Gal4)转录调控下Gal4转基因的母系和父系遗传后果,我们证明Gal4表达可能对发育和寿命有害。与在其他Gal4转基因控制下表达UAS-lacZ转基因的果蝇相比,在存在UAS-lacZ转基因的情况下表达arm-Gal4的雄性后代小眼数量和大小减少。在25°C下饲养,母系遗传elav-Gal4且无响应转基因或带有UAS-lacZ的雄性果蝇的中位寿命为70天。母系遗传arm-Gal4且无响应转基因的雄性果蝇的中位寿命为48天,带有UAS-lacZ转基因时为40天。在父系arm-Gal4控制下表达UAS-lacZ可部分挽救这种表型,平均中位寿命为60天。这些数据表明,arm-Gal4对果蝇发育和寿命具有有害影响,且这种影响直接取决于亲本遗传,并且良性响应和报告基因UAS-lacZ可能会影响黑腹果蝇的发育。在设计和实施UAS-Gal4表达实验时应考虑这些发现。