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!
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
*unsolicited mail is *NOT* appreciated, usual disclaimers apply
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