Re: [ccp4bb] Semet in non-auxotrophic strains

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

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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

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Subject: Re: Semet in non-auxotrophic strains
From: Cynthia Kinsland clk10 {- at -} CORNELL {- dot -} EDU
Date: 2008-11-03

You can express Se-Met protein in non-auxotrophs using metabolic
inhibition (Doublie. Methods Enzymol. 276, 523-530, 1997 and Van Duyne
et al. J. Mol. Biol. 229, 105-124, 1993).

The Studier auto-induction paper (Studier, Prot. Exp. Purif. 41,
207-234, 2005) describes an auto-induction Se-Met medium that gives
sufficient incorporation of Se-Met for phasing in non-auxotrophs.

Good luck,

Cynthia

On Nov 3, 2008, at 12:56 PM, Jacob Keller wrote:

> 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
> *******************************************

____________________
Cynthia Kinsland, Ph.D.
Cornell University
Protein Facility Director
607-255-8844

CCP4bb navigation

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

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

Next message:
Subject: Re: Cryoprotectant for protein-DNA complex crystal
From: Phoebe Rice price {- at -} UCHICAGO {- dot -} EDU
Date: 2008-11-03