Expression of peptides in E.Coli

Koen De Smet k.desmet at nospam.ic.ac.uk
Mon Jun 15 02:40:19 EST 1998


Svend Kjaer wrote:
> 
> Hi all,
> 
> I am in need of a peptide of the length of roughly 20 amino acids. Instead of
> having someone to synthesize it (who I will then have to pay), I thought of
> expressing it into the periplasme of E.coli and purifying it via a His-tag.
> However, small non-structured peptides of that length may be degraded by pro
> teases, so I would like to know if anyone has experience with such set-ups
> 
> Thanks,
> 
> Svend Kjaer,
> 

I did something similar a few years ago (De Smet et al, J. Immunol. Meth. 177, 243-250, 
1994). We used the pGEX-2T expression vector, which gives a fusion protein with the 
20-40 aa peptide at the end. These could be bound on glutatione-affinity columns and 
the peptide cleaved off with thrombin. It worked, but we decided to continue making 
peptides by chemical synthesis. The latter gives you more peptide, unless you wantto go 
to large fermenters.


Koen


Koen De Smet
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