I'm looking for a way to prevent fluorescence of GFP after translation, which can be reversed quickly using some post-translational modification. Ideally translated GFP would be inactive until phosphorylation (or something similar), at which point it begins to fluoresce.
Hi David,
I only know about pH sensitive GFP variants - used to study exocytosis and endocytosis. E.g. in acidic pH GFP does not show fluorescence and in neutral pH the GFU has fluorescence.
If you think that this could be an option, I can tell you about those molecules.
Hi,
You have avaiable this construction Ub-G76V-GFP (https://www.addgene.org/11941/). The fusion protein is constantly degraded in the UPS (you should see some low-to-basal fluorescence). However, in order to prevent the degradation, you should, at least, mildly inhibit the proteasome. Not sure if it was helpful.
Good luck
- Badges
- Science topic
- Similar topics
- Physical Sciences
- Physical Phenomena
- Luminescence
- Fluorescence