Help: GST-protein cleaved.
D.K.
no.email at thanks.to.spam.net
Sun Apr 13 14:18:39 EST 2003
In article <20030407203914.6267.qmail at ww02.hostica.com>, hyi at ksu.edu wrote:
>
>I am expressing a GST-protein (vector=pGEX-2T, host=BL-21), but the product is
> cleaved so I have two populations of expressed protein. Can anybody tell me
> how to solve this problem ? Thank you very much.
Nothing except the generic advice is possible:
1) Find out if the cleavage is in vivo or in vitro. If it vitro, use
proper protease inhibitors to suppress. If in vivo then
2) change host and or/vector/fusion partner and see how you fair.
3) If everything else fails, purify one from another using either
i) conventional approaches or ii) by introducing second tag
on C-terminus.
DK
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