Tau-β-Galactosidase, an Axon-Targeted Fusion Protein
The most commonly used enzymatic reporter molecule, Escherichia coli β-galactosidase (β-gal; β-D-galactoside galactohydrolase, EC 3.2.1.23), fails to readily diffuse into axons; consequently, the morphologies of β-gal-labeled neurons cannot directly be determined. For analysis of neuronal pathfindin...
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Veröffentlicht in: | Proceedings of the National Academy of Sciences - PNAS 1994-06, Vol.91 (13), p.5972-5976 |
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Sprache: | eng |
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Zusammenfassung: | The most commonly used enzymatic reporter molecule, Escherichia coli β-galactosidase (β-gal; β-D-galactoside galactohydrolase, EC 3.2.1.23), fails to readily diffuse into axons; consequently, the morphologies of β-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 β-gal reporter by fusing the cDNA encoding the bovine microtubule-binding protein, tau, to lacZ, the E. coli gene encoding β-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-β-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 β-gal in vertebrate neurons and prove to be a useful reagent for the analysis of vertebrate nervous systems. |
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ISSN: | 0027-8424 1091-6490 |
DOI: | 10.1073/pnas.91.13.5972 |