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