[ccp4bb] protein expression problem

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CCP4bb <-- 2008 <-- January 2008 <-- 22 January 2008

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Subject: Re: CCP4 for bioinformatics?
From: Gerard DVD Kleywegt gerard {- at -} XRAY {- dot -} BMC {- dot -} UU {- dot -} SE
Date: 2008-01-21

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Subject: Re: protein expression problem
From: David Briggs drdavidcbriggs {- at -} GMAIL {- dot -} COM
Date: 2008-01-22

Subject: protein expression problem
From: Daniel Jin jin_xiechen {- at -} YAHOO {- dot -} COM
Date: 2008-01-22

Hi,

I have been trying to express a rat protein in bacteria. The MBP-fusion expressed at very high level (~ 40 mg/L) while the GST-fusion and His-tag only gave inclusion bodies. The problem is that all protein runs in the void volume on a size-exclusion column (s-200, hepes, pH 7.4, 200 mM NaCl), no matter it is the intact MBP-fusion or cleaved sample. There is no Cys on this protein so there is unlikely any disulfide bond related problem. Anything I can do before I throw away this construct and try insect or mammalian cells? Thanks.

Best,
Chen

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CCP4bb navigation

CCP4bb <-- 2008 <-- January 2008 <-- 22 January 2008

Previous message:
Subject: Re: CCP4 for bioinformatics?
From: Gerard DVD Kleywegt gerard {- at -} XRAY {- dot -} BMC {- dot -} UU {- dot -} SE
Date: 2008-01-21

Next message:
Subject: Re: protein expression problem
From: David Briggs drdavidcbriggs {- at -} GMAIL {- dot -} COM
Date: 2008-01-22