Clonetech 'Matchmaker' vectors
cgdn at uhura.cc.rochester.edu
Fri Sep 29 23:31:02 EST 1995
Clonetech (sic) Matchmaker vectors are basically the system developed
by Stephen Elledge. The fusion protein expression is generally high
enough to easily be detected by Western blotting. Clontech will sell
you monoclonal antibody to the GAL4 activation domain, so you don't
even have to have an Ab to the protein domain you are fusing.
Expression of the fusion protein is under the strong ADH promoter
and the vector is a 2-micron circle which is present in many copies/cell
so the only thing which would lead to low expression levels is
instability of the fusion protein. In my experience the opposite is
the problem. Many of the protein domains I have tried are toxic to
the yeast cells when incorporated in the fusion proteins.
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