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pMal-c2, modification of

Wolfgang Schechinger Wolfgang.Schechinger at med.uni-tuebingen.de
Thu Jan 27 13:40:32 EST 2000

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!


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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