Rapidly degrading GFP-fusion proteins

Details Rapidly degrading GFP-fusion proteins Rapidly degrading GFP-fusion proteins

1998-04-17

2000-10-10

Eyler, Yvonne

Chemistry: natural resins or derivatives; peptides or proteins;

Peptides of 3 to 100 amino acid residues

25 or more amino acid residues in defined sequence

435 77, 436546, 436800, A61K 3800, C07K 500, C07K 700, C07K 1600, C07K 1700

Patent

active

061303135

ABSTRACT:
Green fluorescent protein (GFP) is widely used as a reporter in determining gene expression and protein localization. The present invention provides fusion proteins with a half life of ten hours or less. Such proteins may be constructed by fusing C-terminal amino acids of the degradation domain of mouse ornithine decarboxylase (MODC), which contains a PEST sequence, to the C-terminal end of an enhanced variant of GFP (EGFP). Fluorescence intensity of the fusion protein in transfected cells is similar to that of EGFP, but the fusion protein, unlike EGFP, is unstable in the presence of cycloheximide. Specific mutations in the MODC region have resulted in mutants with varying half lives, useful for a variety of purposes.

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