[ccp4bb] GST fusion-Expression problem

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CCP4bb <-- 1999 <-- November 1999 <-- 30 November 1999

Previous message:
Subject: Re: high tempterature factor refinement problem
From: "Francisco J {- dot -} Enguita" fenguita {- at -} ITQB {- dot -} UNL {- dot -} PT
Date: 2008-10-17

Next message:
Subject: Re: GST fusion-Expression problem
From: "Mark J {- dot -} van Raaij" mark {- dot -} vanraaij {- at -} USC {- dot -} ES
Date: 2008-10-17

Subject: GST fusion-Expression problem
From: "Putcha, Balananda Dhurjati Kumar" bputcha {- at -} UTK {- dot -} EDU
Date: 2008-10-17

Dear CCP4ers,

I have a GST fusion protein (eukaryotic). When I express it in E.coli BL21 (DE3) or RIPL cells, however, I mostly get GST and very little fusion protein. I get very little "leaky expression". I normally grow my cells at 37C in LB or 2X YT and after induction at 20C for 16-18hrs. I also have tried growing at 37C for 2hrs (after induction) with little change. Is there a trick to solving this problem? Thank you for your thoughts
Kumar

CCP4bb navigation

CCP4bb <-- 1999 <-- November 1999 <-- 30 November 1999

Previous message:
Subject: Re: high tempterature factor refinement problem
From: "Francisco J {- dot -} Enguita" fenguita {- at -} ITQB {- dot -} UNL {- dot -} PT
Date: 2008-10-17

Next message:
Subject: Re: GST fusion-Expression problem
From: "Mark J {- dot -} van Raaij" mark {- dot -} vanraaij {- at -} USC {- dot -} ES
Date: 2008-10-17