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Re: [ccp4bb]: Se-met protein



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You might use glycerol as carbon source instead of glucose, even cell
growth is slow, but no inhibition observed. Your induction might also need
to be optimized again.

-------------------------------------------------
Lei Zheng

Dept.Chemistry and biochemistry
Univ. Bern
Freiestr.3  3012 Bern
Switzerland

Tel Number: 0041-31-6314370/34 
Fax Number: 0041-31-6318057
Email: zheng.lei@ioc.unibe.ch
-------------------------------------------------




On Thu, 23 Jan 2003, 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
> 
> **********************************************
> 
>