TAU-BETA-GALACTOSIDASE, AN AXON-TARGETED FUSION PROTEIN
TAU-BETA-GALACTOSIDASE, AN AXON-TARGETED FUSION PROTEIN
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DOI：
10.1073/pnas.91.13.5972
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发表时间：
1994-06-21
影响因子：
11.1
通讯作者：
THOMAS, JB
中科院分区：
文献类型：
--
作者：
CALLAHAN, CA;THOMAS, JB
最常用的酶报告分子，大肠杆菌β-半乳糖苷酶（β-gal; β-D-半乳糖苷半乳糖水解酶，EC 3.2.1.23），不能容易地扩散到轴突;因此，β-gal标记的神经元的形态不能直接确定。对于神经元寻路和突触连接的分析，这些信息是必不可少的。我们通过将编码牛微管结合蛋白tau的cDNA与lacZ融合，构建了一个轴突靶向的β-gal报告基因。coli中编码β-gal的基因。当发育和成年果蝇神经元表达时，该报告者标记细胞体和轴突。它还揭示了非神经元细胞的整个细胞范围，如肌纤维和神经胶质。为了产生用于研究果蝇神经发育的神经元标记物，我们构建了一个tau-β-gal增强子-陷阱转座子。从1500个独立的线产生的动员这种转座子，我们已经分离出一组有用的标记的特定子集的神经元，神经胶质细胞和肌肉。由于tau cDNA-lacZ报告基因利用牛tau蛋白，它也可以有效地靶向脊椎动物神经元中的β-gal，并证明是用于脊椎动物神经系统分析的有用试剂。
The most commonly used enzymatic reporter molecule, Escherichia coli beta-galactosidase (beta-gal; beta-D-galactoside galactohydrolase, EC 3.2.1.23), fails to readily diffuse into axons; consequently, the morphologies of beta gal-labeled neurons cannot directly be determined. For analysis of neuronal pathfinding and synaptic connectivity, this information is essential. We have constructed an axon-targeted beta-gal reporter by fusing the cDNA encoding the bovine microtubule-binding protein, tau, to lacZ, the E. coli gene encoding beta-gal. This reporter labels cell bodies and axons when expressed by developing and adult Drosophila neurons. It also reveals the entire cellular extent of nonneuronal cells such as muscle fibers and glia. To generate neuronal markers for studies of Drosophila neural development, we constructed a tau-beta-gal enhancer-trap transposon. From 1500 independent lines generated by mobilization of this transposon, we have isolated a set of useful markers for specific subsets of neurons, glia, and muscles. Since the tau cDNA-lacZ reporter utilizes bovine tau, it may also effectively target beta-gal in vertebrate neurons and prove to be a useful reagent for the analysis of vertebrate nervous systems.