Re: [ccp4bb] Semet in non-auxotrophic strains

- Protein crystallography

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- Crystallisation

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

Previous message:
Subject: Semet in non-auxotrophic strains
From: Jacob Keller j-keller2 {- at -} MD {- dot -} NORTHWESTERN {- dot -} EDU
Date: 2008-11-03

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Subject: Re: Semet in non-auxotrophic strains
From: Arthur Laganowsky art {- at -} MBI {- dot -} UCLA {- dot -} EDU
Date: 2008-11-03

Subject: Re: Semet in non-auxotrophic strains
From: Guenter Fritz guenter {- dot -} fritz {- at -} UNI-KONSTANZ {- dot -} DE
Date: 2008-11-03

Hi Jacob,
you can express SeMet labeled protein in almost any E.coli expression
strain if you use a defined minimal medium.
Here is a receipe that worked quite well in our hands:
http://strucbio.biologie.uni-konstanz.de/ccp4wiki/index.php/Expression_of_SeMet_labeled_proteins
HTH
Guenter
> Dear Crystallographers,
>
> I have been using an E coli strain called Tuner (DE3) pLacI, which is
> not auxotrophic for methionine, nor is there an auxotrophic analogue
> available. It may be that my protein can be expressed in other
> strains, but this one works well right now, and it is usually better
> to stay with what works well. Has anybody tried growing semet protein
> in non-auxotrophic strains, or are there tricks to using
> non-auxotrophs for semet protein?
>
> Thanks,
>
> Jacob
>
> *******************************************
> Jacob Pearson Keller
> Northwestern University
> Medical Scientist Training Program
> Dallos Laboratory
> F. Searle 1-240
> 2240 Campus Drive
> Evanston IL 60208
> lab: 847.491.2438
> cel: 773.608.9185
> email: j-keller2@northwestern.edu
> *******************************************

--
***********************************

Priv.Doz.Dr. Guenter Fritz
Fachbereich Biologie
Sektion Naturwissenschaften
Universitaet Konstanz
http://www.biologie.uni-konstanz.de/fritz

Universitaetsstrasse 10
Postfach M665
D-78457 Konstanz

e-mail: Guenter.Fritz@uni-konstanz.de

Tel. Office: +49-(0)7531 88 3205
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CCP4bb navigation

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

Previous message:
Subject: Semet in non-auxotrophic strains
From: Jacob Keller j-keller2 {- at -} MD {- dot -} NORTHWESTERN {- dot -} EDU
Date: 2008-11-03

Next message:
Subject: Re: Semet in non-auxotrophic strains
From: Arthur Laganowsky art {- at -} MBI {- dot -} UCLA {- dot -} EDU
Date: 2008-11-03