peptide expression in E.coli

Ben Davis bjd12 at cus.cam.ac.uk
Fri Mar 24 11:24:39 EST 1995


M.R.douglas at rheuma.bham.ac.uk wrote:
: I am interested in producing peptides (10-30 a.a. in length) in E.coli with a
: view to labelling them isotopically for NMR experiments.  Does anyone out
: there have any hints, tips or references as to the best way to go about this?
: And which is the best purification method for sorting out the good from the bad?
: Getting them synthesised in the old fashioned way would break the bank, I'm
: afraid.

Hi

My guess is that you've got basically no chance of expressing anything that
small; coli proteases will chew it up & spit it out

I'd recommend getting a small protein to fuse it onto, and overexpressing
that fusion protein, then cleave it off. You waste the labelling that goes
into the other protein, but 15N is (relatively) cheap, and as far as I can
see its the only way of getting labelled bits the size you want.

(I work on labelled 40 & 20 mers btw, but make them by cleaving a 60 residue
protein)

: Here's hoping!

Good luck

: Mike.

Ben

--
______________________________________________________________________________

Ben Davis,
MRC Protein Function and Design,
Cambridge, UK
______________________________________________________________________________

"They can make me do it, but they can't make me do it with dignity."



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