Re: [ccp4bb] Inclusion body

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

Previous message:
Subject: Re: Inclusion body
From: Artem Evdokimov artem {- at -} XTALS {- dot -} ORG
Date: 2007-03-23

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Subject: Re: Expression hosts (was:Inclusion body)
From: Dima Klenchin klenchin {- at -} FACSTAFF {- dot -} WISC {- dot -} EDU
Date: 2007-03-23

Subject: Re: Inclusion body
From: Ashley Buckle ashley {- dot -} buckle {- at -} MED {- dot -} MONASH {- dot -} EDU {- dot -} AU
Date: 2007-03-23

Dear Weijun

Have a look at our REFOLD database - there are some examples of
chaperone assisted refolding as well as many protocols for refolding
of disulphide-rich proteins

http://refold.med.monash.edu.au

good luck
ashley

On 24/03/2007, at 12:23 PM, verdino@SCRIPPS.EDU wrote:

> Hi Weijun,
>
> you can get a kit from Takara (chaperone kit #3340) which has five
> different chaperone plasmids (combinations of various chaperone). It
> worked for me assofar that I could express huge amounts of the
> chaperones,
> but my target protein was either not expressed at all any more
> (seems as
> if E. coli was all busy expressing chaperons) or still insoluble ;-(
> And unfortunately the structures of the chaperones were all solved
> already
> ;-)
>
> If disulfides might be the problem - try expression into the
> periplasma
> (some pET vectors encode leader sequences for that).
>
> Otherwise try other expression systems (baculovirus, mammalian....)
>
> All the best, Petra
>
>
>> Dear All,
>>
>> I got enclusion body in many cases when I tried to express human
>> proteins in E coli. I would like some suggestions on how to go about
>> it. I would also like to try co-expression of GroEL/GroES or
>> DnaJ/DnaK, and would like to know where to get the plasmids.
>>
>> Any help or comments would be appreciated.
>> Weijun
>>
>
>
> --
> Petra Verdino, PhD
> Research Associate
> The Scripps Research Institute, BCC206
> 10550 North Torrey Pines Road
> CA 92037 La Jolla, USA
> tel:1-858-784-2294
> fax:1-858-784-2980

Ashley Buckle Ph.D
NHMRC Senior Research Fellow
The Department of Biochemistry and Molecular Biology
School of Biomedical Sciences, Faculty of Medicine &
Victorian Bioinformatics Consortium (VBC)
Monash University, Clayton, Vic 3800
Australia

http://www.med.monash.edu.au/biochem/staff/abuckle.html
iChat/AIM: blindcaptaincat
skype: ashley.buckle
Tel: (613) 9905 3781 (office)
Tel: (613) 9905 1653 (lab)

Fax : (613) 9905 4699

CCP4bb navigation

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

Previous message:
Subject: Re: Inclusion body
From: Artem Evdokimov artem {- at -} XTALS {- dot -} ORG
Date: 2007-03-23

Next message:
Subject: Re: Expression hosts (was:Inclusion body)
From: Dima Klenchin klenchin {- at -} FACSTAFF {- dot -} WISC {- dot -} EDU
Date: 2007-03-23