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Titolo:
Direct observation of nucleocytoplasmic transport by microinjection of GFP-tagged proteins in living cells
Autore:
Rosorius, O; Heger, P; Stelz, G; Hirschmann, N; Hauber, J; Stauber, RH;
Indirizzi:
Univ Erlangen Nurnberg, Inst Med & Clin Virol, D-91054 Erlangen, Germany Univ Erlangen Nurnberg Erlangen Germany D-91054 -91054 Erlangen, Germany
Titolo Testata:
BIOTECHNIQUES
fascicolo: 2, volume: 27, anno: 1999,
pagine: 350 -
SICI:
0736-6205(199908)27:2<350:DOONTB>2.0.ZU;2-7
Fonte:
ISI
Lingua:
ENG
Soggetto:
NUCLEAR-ENVELOPE; BINDING; EXPORT;
Tipo documento:
Article
Natura:
Periodico
Settore Disciplinare:
Life Sciences
Citazioni:
17
Recensione:
Indirizzi per estratti:
Indirizzo: Stauber, RH Univ Erlangen Nurnberg, Inst Med & Clin Virol, Schlossgarten 4, D-91054 Erlangen, Germany Univ Erlangen Nurnberg Schlossgarten 4 Erlangen Germany D-91054
Citazione:
O. Rosorius et al., "Direct observation of nucleocytoplasmic transport by microinjection of GFP-tagged proteins in living cells", BIOTECHNIQU, 27(2), 1999, pp. 350

Abstract

We established a straightforward experimental system to investigate directly the requirements for nucleocytoplasmic transport in live cells. For thispurpose, 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 res 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, thekinetics of nuclear import/export in living cells and to evaluate qualitative differences in various NLS/NES signals and pathways.

ASDD Area Sistemi Dipartimentali e Documentali, Università di Bologna, Catalogo delle riviste ed altri periodici
Documento generato il 10/07/20 alle ore 03:06:08