Sun B, Xu P, Salvaterra P M
Division of Neuroscience, Beckman Research Institute of the City of Hope, 1450 East Duarte Road, Duarte, CA 91010, USA.
Proc Natl Acad Sci U S A. 1999 Aug 31;96(18):10438-43. doi: 10.1073/pnas.96.18.10438.
We have constructed transgenic Drosophila melanogaster lines that express green fluorescent protein (GFP) exclusively in the nervous system. Expression is controlled with transcriptional regulatory elements present in the 5' flanking DNA of the Drosophila Na(+), K(+)-ATPase beta-subunit gene Nervana2 (Nrv2). This regulatory DNA is fused to the yeast transcriptional activator GAL4, which binds specifically to a sequence motif termed the UAS (upstream activating sequence). Drosophila lines carrying Nrv2-GAL4 transgenes have been genetically recombined with UAS-GFP (S65T) transgenes (Nrv2-GAL4+UAS-GFP) inserted on the same chromosomes. We observe strong nervous system-specific fluorescence in embryos, larvae, pupae, and adults. The GFP fluorescence is sufficiently bright to allow dynamic imaging of the nervous system at all of these developmental stages directly through the cuticle of live Drosophila. These lines provide an unprecedented view of the nervous system in living animals and will be valuable tools for investigating a number of developmental, physiological, and genetic neurobiological problems.
我们构建了转基因黑腹果蝇品系，该品系仅在神经系统中表达绿色荧光蛋白（GFP）。表达由果蝇Na(+)、K(+)-ATP酶β亚基基因Nervana2（Nrv2）的5'侧翼DNA中存在的转录调控元件控制。该调控DNA与酵母转录激活因子GAL4融合，GAL4特异性结合称为UAS（上游激活序列）的序列基序。携带Nrv2-GAL4转基因的果蝇品系已与插入同一染色体上的UAS-GFP（S65T）转基因进行遗传重组（Nrv2-GAL4+UAS-GFP）。我们在胚胎、幼虫、蛹和成虫中观察到强烈的神经系统特异性荧光。GFP荧光足够明亮，可直接通过活果蝇的表皮对所有这些发育阶段的神经系统进行动态成像。这些品系为活体动物的神经系统提供了前所未有的视角，将成为研究许多发育、生理和遗传神经生物学问题的有价值工具。
