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 

More information about the Methods mailing list