pGEX/GST gene fusion problems-> HELP!

Paul N Hengen pnh at ncifcrf.gov
Fri Aug 23 16:52:21 EST 1996


Mark Gosink (mmgosink at facstaff.wisc.edu) wrote:

> While our lab does not use GST fusions, we do express quite a few 
> other fusion proteins. We have found that in addition to using 
> relatively short induction times (2-3hr) that I see others have 
> suggested we also use lower incubation temperatures (i.e. 30C or even 
> ~22C) at times. (Note: If you use lower temperatures you may find that 
> your yield is decreased but the percentage of undegraded protein 
> increases.)

Do you have any idea why this works? Is it that the enzymes or undefined
proteases are less active at this temperature, or is it that they are not
expressed at this temperature?

--
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