Direct observation of nucleocytoplasmic transport by microinjection of GFP-tagged proteins in living cells
We established a straightforward experimental system to investigate directly the requirements for nucleocytoplasmic transport in live cells. For this purpose, substrates were created containing nuclear localization signals (NLS) or nuclear export signals (NES) linked to a chimeric protein composed o...
| Veröffentlicht in: | BioTechniques. - 1993. - 27(1999), 2 vom: 22. Aug., Seite 350-5 |
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| 1. Verfasser: | |
| Weitere Verfasser: | , , , , |
| Format: | Aufsatz |
| Sprache: | English |
| Veröffentlicht: |
1999
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| Zugriff auf das übergeordnete Werk: | BioTechniques |
| Schlagworte: | Journal Article Research Support, Non-U.S. Gov't Luminescent Proteins Nuclear Localization Signals Green Fluorescent Proteins 147336-22-9 Glutathione Transferase EC 2.5.1.18 |
| Zusammenfassung: | We established a straightforward experimental system to investigate directly the requirements for nucleocytoplasmic transport in live cells. For this purpose, substrates were created containing nuclear localization signals (NLS) or nuclear export signals (NES) linked to a chimeric protein composed of the glutathione S-transferase (GST) fused to the green fluorescent protein (GFP). The combination of GST/GFP-tagging allowed us to control protein expression in bacteria and to monitor protein purification during chromatography. Following microinjection into somatic cells, nuclear export/import of the highly fluorescent substrates could be observed directly by fluorescence microscopy. This system sets the stage to quantitate, in real time, the kinetics of nuclear import/export in living cells and to evaluate qualitative differences in various NLS/NES signals and pathways |
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| Beschreibung: | Date Completed 20.09.1999 Date Revised 10.12.2019 published: Print Citation Status MEDLINE |
| ISSN: | 0736-6205 |