de-stabilized GFP

Model Organism: Worms Scientific Discussion
1

I am hoping to generate a GFP with a shortened half-life for expression in worms. Has anyone experimented with this?

I 've read that a “PEST” sequence (from ornithine decarboxylase) has been used in mammalian systems to decrease GFP stability, but the worm enzyme lacks this domain (naturally). Does anyone know if this (i.e., PEST-mediated protein degradation) might work in worms? Some proteins do seem to have PEST sequences, but I can’t figure out if they have been demonstrated to actually enhance protein turnover in those particular worm proteins.

Alternatively, any other strategy to shorten GFP half-life in worms would be valuable. Please post suggestions, even if half-baked.

2

see…“Frand AR, Russel S, Ruvkun G.
Functional genomic analysis of C. elegans molting.
PLoS Biol. 2005 Oct;3(10):e312”…they used the PEST (Pro-Glu-Ser-Thr) to decrease turnover to 1hr in vivo

3

Thank you! I hadn’t yet come across that reference, and it’s exactly what I was trying to find out. Much appreciated.