HIS-tagged Protein Purification/Ni-NTA-Agarose

Tom Thatcher ttha at troi.cc.rochester.edu
Wed Dec 15 08:32:45 EST 1993


In <2eli4r$67v at news.u.washington.edu> findley at carson.u.washington.edu (Seth Findley) writes:

>Does anybody have any further advise on overcoming degradation PRIOR to
>the purification steps?
>(ie: optimization of growth conditions in E.coli host)

You should know that some proteins are just unstable in E. coli
no matter what you do.  A postdoc in our lab was expressing
deletion clones of a particular protein.  A 10 amino acid C-terminal
deletion was expressed at high levels, but the full length protein
was not expressed, or was degraded, or was toxic to the host.

Tom Thatcher
--
Tom Thatcher
University of Rochester
School of Medicine and Dentistry
ttha at troi.cc.rochester.edu 



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