Hi
I am trying to fuse 2 proteins and express them in mammalian cells. One of the proteins is a membrane receptor and the other is a soluble protein. I have to create 2 stable cell lines, one expressing the fusion protein and the other expressing only the soluble protein. However, I am interested in creating a system where I have to create only one stable cell line with the fusion protein containing a region between them which I can inducibly cleave using small molecule or proteases, etc.
I would really appreciate if anyone could direct me in the right direction and kindly provide me with references so I can look up how efficient that technique is.
Regards
Viruses do this all the time, so in theory you could transfect a viral protease to cleave your fusion protein with the correct cleavage sequence, however these proteases cleave many host factors as well and differences you may be detecting can be artefactual. There is a system that can be used to do this that is used in yeast and some mammalian cells. it's called the iDimerize system where you fuse your genes of interest to Dmr domains and then adding a compound similar to rapamycin to the culture media causes these two proteins to dimerize, but you have to control properly for the drug alone as it still can affect mTOR (though its makers claim it's inactive). Here's the link if you want to check it out.
http://www.clontech.com/US/Products/Inducible_Systems/Protein-Protein_Interactions/Homodimer
Good luck!
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