Nuclear import in digitonin-permeabilized cells.
Import of proteins into the nucleus of a cell is a complex process that can be reconstituted in vitro. Digitonin-permeabilized cells are washed free of cytosolic factors to provide competent nuclei. Cytosolic factors for import are provided by an extract of Xenopus ovarian cells. Fluorochrome-conjugated probes, cloned proteins fused to green fluorescent protein, or antibodies to the protein of interest are used to visualize nuclear import. The system can also be used to identify nuclear localization sequences (NLS) of imported proteins.
Moore
M S
MS
Baylor College of Medicine, Houston, Texas, USA.
Schwoebel
E D
ED
D021581:Active Transport, Cell Nucleus / Q000502:physiology*; D000818:Animals; D002463:Cell Membrane Permeability / Q000187:drug effects / Q000502:physiology; D003584:Cytological Techniques; D004072:Digitonin / Q000494:pharmacology*; D005260:Female; D006367:HeLa Cells; D006801:Humans; D007202:Indicators and Reagents / Q000494:pharmacology*; D010053:Ovary / Q000166:cytology*; D014981:Xenopus
D007202:Indicators and Reagents; D004072:Digitonin