IUBio Biosequences .. Software .. Molbio soft .. Network News .. FTP

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!

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
-----
*unsolicited mail is *NOT* appreciated, usual disclaimers apply 
-----                         
Pressing ALT+F4 simultaneously enables you to read private messages
---




More information about the Methods mailing list

Send comments to us at biosci-help [At] net.bio.net