Ubiquitin-fusion protein system: a powerful tool for ectopic protein expression in mammalian cells.
Clonal cell lines stably expressing a protein of interest are frequently used in the characterization of the potential physiological functions of mammalian proteins, but the generation of such cell lines is laborious and time-consuming. Here, we made use of the fact that fusion proteins consisting of ubiquitin linked to the N terminus of a protein of interest are efficiently processed by ubiquitin-specific proteases into their respective free proteins within eukaryotic cells, and designed an expression system that allows easy and highly efficient selection of mammalian cells ectopically expressing a protein of interest.
Matentzoglu
Konstantin
K
University of Konstanz, Konstanz, Germany.
Scheffner
Martin
M
D002460:Cell Line; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism; D006801:Humans; D015202:Protein Engineering / Q000379:methods*; D011993:Recombinant Fusion Proteins / Q000378:metabolism*; D014162:Transfection; D025801:Ubiquitin / Q000235:genetics* / Q000378:metabolism; D044767:Ubiquitin-Protein Ligases / Q000235:genetics / Q000378:metabolism
D011993:Recombinant Fusion Proteins; D025801:Ubiquitin; C485184:enhanced green fluorescent protein; D049452:Green Fluorescent Proteins; C471639:UBE3A protein, human; D044767:Ubiquitin-Protein Ligases