Attenuation of green fluorescent protein half-life in mammalian cells

Protein Eng. 1999 Dec;12(12):1035-40. doi: 10.1093/protein/12.12.1035.


The half-life of the green fluorescent protein (GFP) was determined biochemically in cultured mouse LA-9 cells. The wild-type protein was found to be stable with a half-life of approximately 26 h, but could be destabilized by the addition of putative proteolytic signal sequences derived from proteins with shorter half-lives. A C-terminal fusion of a PEST sequence from the mouse ornithine decarboxylase gene reduced the half-life to 9.8 h, resulting in a GFP variant suitable for the study of dynamic cellular processes. In an N-terminal fusion containing the mouse cyclin B1 destruction box, it was reduced to 5.8 h, with most degradation taking place at metaphase. The combination of both sequences produced a similar GFP half-life of 5.5 h. Thus, the stability of this marker protein can be controlled in predetermined ways by addition of the appropriate proteolytic signals.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Cell Line
  • Cyclin B / genetics
  • Cyclin B1
  • Flow Cytometry
  • Fluorescence
  • Green Fluorescent Proteins
  • Kinetics
  • Luminescent Proteins / metabolism*
  • Mice
  • Ornithine Decarboxylase / genetics
  • Protein Sorting Signals / genetics
  • Recombinant Fusion Proteins / metabolism
  • S Phase
  • Transfection


  • Ccnb1 protein, mouse
  • Cyclin B
  • Cyclin B1
  • Luminescent Proteins
  • Protein Sorting Signals
  • Recombinant Fusion Proteins
  • Green Fluorescent Proteins
  • Ornithine Decarboxylase