Hi all,
I would like to express a His-tagged protein in E. coli. The only
suitable vector I have at the moment, is pMal-c2.
There is a unique restriction site (NdeI) right
between the Ptac promotor and the start of malE translation.
Thus I would cut out the whole maltose binding protein (and get rid
of it since I don't need it anyway, because I have a His-tag) and
insert my construct between this NdeI site and a the polylinker.
Despite all this sounds quite reasonable to me, does anybody have
any caveats, drawbacks or warnings to mention?
All input is welcome!
Wolfgang
For those who don't know pMal-c2:
pMal-c2 is a glucose inducible vector for procaryotic protein
expression. When used normally, one has a fusion protein between
maltose binding protein (malE) and the protein on interest; the
bacterial lysate is purified by affinity purification using a amylose
(maltose-like polymer) resin. Then the fusion protein is cleaved off
proteolytically with factor Xa.
Dr. Wolfgang Schechinger, Pathobiochemistry Dept.
University of Tuebingen, Germany
email: wolfgang.schechinger at med.uni-tuebingen.de
wwWait: http://www.medizin.uni-tuebingen.de/~wgschech/start.htm
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