proteolytic degradation of rcombinant protein

Vladimir Svetlov svetlov at oncology.wisc.edu
Mon Nov 30 16:08:33 EST 1998


In article <366300B7.E85 at nospam.ipk-gatersleben.de>,
boernke at nospam.ipk-gatersleben.de wrote:


> I have this protein expressed as a MBP-fusion in E. coli XL-I blue
> using NEB's pMAL vector. Unfortunately I only see a faint band in a
> commassie stained gel. However, western blottig shows me several bands
> all smaller than my fusion protein which to me seems like proteolytic
> degradation. Is there any way to avoid this, like lowering temperature
> or changing host strain.
> Any hints will be appreciated.

There is a little review on dealing with protein degradation/fragmentation
in E. coli with a number of pointers to methods papers:

Murby M, Uhlen M, Stahl S
Upstream strategies to minimize proteolytic degradation upon recombinant
production
in Escherichia coli.
Protein Expr Purif 1996 Mar;7(2):129-36.

Cheers,
V.

-- 
Vladimir Svetlov, PhD
McArdle Lab for Cancer Res.
UW-Madison
1400 University Ave.
Madison, WI 53706



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