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Re: [ccp4bb]: Se-met protein
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Dear Kostas :
If you have no other choice that pBAD vector, my suggestion will be to
use a complex medium supplemented with Se-Met (insted of the minimal
one) to obtain an Se-enriched protein. Sometimes it works.
Other possible choice could involve the screening of other carbon
sources for the minimal medium. I don't know what strain are you using
for expression ..
Best regards
Francisco
On Thu, 2003-01-23 at 11:55, Konstantinos Beis wrote:
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>
> Dear All,
> I have to grow selenomethionine derivative protein for MAD collection. I
> have my gene in a pBAD vector and it is arabinose inducible. Unfortunately
> I cannot follow the SeMet incorporation by metabolic inhibition (VanDuyne
> G.D. et. al. (1993) LJM, 229, 105-124) due to the fact that the minimal
> media have glucose as the carbon source, and this will repress the
> expression of the gene from the ara promotor. I can only use the pBAD
> system for my protein. Is there any other way/modification of the above
> protocol that I could grow Se-met protein in the pBAD vector? Any
> suggestions are welcomed.
>
> Thanks,
> Kostas
>
>
> **********************************************
> Konstantinos Beis
> Centre for Biomolecular Sciences
> University of St Andrews
> North Haugh
> St Andrews
> FIFE KY16 9ST
>
>
> tel: +44 1334 427265
> email: kb11@st-andrews.ac.uk
>
> **********************************************
>
--
============================================
Francisco J. Enguita, Ph.D.
Protein Crystallography laboratory
Instituto de Tecnologia Química e Biológica
2781-901 Oeiras
PORTUGAL
Phone : 351-21-4469669
Fax : 351-21-4433644
E-mail : fenguita@itqb.unl.pt
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