Fellow two-hybrid screeners,
I am about to do a two-hybrid screen. Since my polypeptide of interest is
naturally at the N-terminus, I am interested in expressing it in front of the
GAL4 DNA binding domain (DBD) as well as C-terminal to it. I would appreciate
very much hearing from anyone who has made constructs that would be useful for
cloning a bait protein N-terminal to the GAL4 DBD.
Since my minimum requirement is simply to make the N-terminal fusion, a useful
plasmid therefore need not even be of a two-hybrid type; I could subsequently
subclone the bait-GAL4 DBD fusion into the vector I'm already using.