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Övervakning av ubiquitin-proteasom aktivitet i levande celler Använda en Degron (DGN)-destabiliseras grönt fluorescerande protein (GFP)-baserad Reporter protein
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Övervakning av ubiquitin-proteasom aktivitet i levande celler Använda en Degron (DGN)-destabiliseras grönt fluorescerande protein (GFP)-baserad Reporter protein

Article DOI: 10.3791/3327-v • 10:25 min • November 10th, 2012
November 10th, 2012 •
1, 1, 1, 2, 1
1Molecular and Cell Biology, Institute for Biomedical Aging Research, 2Department of Gerontology and Geriatrics, Netherlands Consortium for Healthy Aging, Leiden University Medical Center

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En metod för att övervaka ubiquitin-proteasom aktivitet i levande celler beskrivs. En degron-destabiliserade GFP-(GFP-DGN) och en stabil GFP-dgnFS fusionsprotein genereras och transduceras in i cellen med användning av en Lentiviral expressionsvektor. Denna teknik gör det möjligt att generera en stabil GFP-dgn/GFP-dgnFS uttryckande cellinje i vilken ubiquitin-proteasom aktivitet kan enkelt bedömas med epifluorescens eller flödescytometri.

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Cellbiologi molekylärbiologi medicin medicinsk teknik virologi proteasom aktivitet lentivirala partiklar GFP-DGN GFP-dgnFS GFP humana diploida fibroblaster flödescytometri plasmid vektor
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