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Hi everyone,
I have a question about doing yeast two hybridization for proteins from C. elegans.
I have checked several papers and all of them did the Y2H at 30 degree.
Is it better to do it at lower temperature such as 25 degree to allow correct folding of worm protein,
Since C. elegans normally grow at 20-25 degree, and we usually express worm proteins at low temperature such as 16 degree?
many thanks,
Ye
An obvious option would be to adapt yeast two-hybrid to another, more cryophilic yeast. Kudriavzevii, for example, the species that was used to provide a version of Gal4p that would work better at C. elegans cultivation temperatures. But you might have trouble using cerevisiae, because it doesn’t thrive at the temperatures at which we cultivate C. elegans.
We always used 30 deg for this and it worked well for screens of elegans libraries and pairwise tests. You are right though that the high temp could be an issue for some worm proteins. Typical yeast strains for 2-hybrid will still grow at 25 deg, albeit more slowly, so you could try that too.
As a part of migration from the Worm Community Forum, this post was manually created on behalf of the user YeHong; when they join the forum here, they can claim this post by replying to this topic.
Thanks for your replies and suggestions. I have tried 25 degree using PJ69-4A strain and it works well.