Refining GAL4-driven transgene expression in Drosophila with a GAL80 enhancer-trap
Refining GAL4-driven transgene expression in Drosophila with a GAL80 enhancer-trap
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DOI：
10.1002/gene.20051
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发表时间：
2004-08-01
期刊：
影响因子：
1.5
通讯作者：
Sokolowski, MB
中科院分区：
文献类型：
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作者：
Suster, ML;Seugnet, L;Sokolowski, MB
我们构建了一个增强子陷阱元件，P {GAL80}，它编码酵母GAL80阻遏物，以改善由二元GAL4/UAS系统驱动的转基因表达。GAL80通过结合其转录激活结构域来阻断GAL4活性。我们在完整的幼虫神经系统中筛选了抑制GAL4诱导的绿色荧光蛋白（GFP）的GAL80增强子陷阱。我们选择了一个线，抑制绿色荧光蛋白在一个大的胆碱能神经元。该细胞系用于将GFP表达从一组超过200个神经元细化到预选GAL4细胞系中的20个神经元的子集。破伤风神经毒素，神经递质释放的一种有效的阻滞剂，在这20个神经元的表达再现了一个异常的幼虫转向行为先前分配给父母的200个神经元。我们的研究结果表明，有针对性的GAL80的表达可能成为一个有用的手段空间精炼转基因表达在果蝇。(C)2004 Wiley-Liss，Inc.
We constructed an enhancer-trap element, P{GAL80}, that encodes the yeast GAL80 repressor to refine expression of transgenes driven by the binary GAL4/UAS system. GAL80 blocks GAL4 activity by binding to its transcriptional activation domain. We screened GAL80 enhancer-traps for repression of GAL4-induced green fluorescent protein (GFP) in the intact larval nervous system. We selected one line that repressed GFP in a large set of cholinergic neurons. This line was used to refine GFP expression from a set of over 200 neurons to a subset of 20 neurons in a preselected GAL4 line. Expression of tetanus neurotoxin, a potent blocker of neurotransmitter release, in these 20 neurons reproduced an aberrant larval turning behavior previously assigned to the parental set of 200 neurons. Our results suggest that targeted GAL80 expression could become a useful means of spatially refining transgene expression in Drosophila. (C) 2004 Wiley-Liss, Inc.