Hi :)
In addition to excellent replies already posted, a few thoughts:
1. Do the whole-protein MS of your stuff. Assuming that you get a spectrum then
a) If you are right and it is indeed your recombinant GST - you
will see a mass pattern that's interpretable via analysis of probable
cleavage
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- Reply message -
From: "Sebastiano Pasqualato"
Date: Fri, Mar 16, 2012 10:12 am
Subject: [ccp4bb] GST-fusion protein production in insect cells
To:
Dear Imre, dear Katya,
thanks a lot for the prompt and insightful replies.
@Imre: well, I don't think it is a matter of linker of lack of Met/presence of
stop codon after GST.
We use the exact same linker in bacteria and that works just nice, while wrt to
the Met, we have cases in which we have
Dear CCP4ers, Dr. Berger,
we have an accumulating series of GST-fusion proteins here that are all
displaying the same behavior when expressed in Hi5 insect cells with the
MultiBac system.
What we are experiencing is a massive production of free GST and only a limited
amount of fusion protein.
