atutter at jeeves.ucsd.edu
Thu Dec 12 23:03:42 EST 1996
Has anyone out there used any expression vector which fuses a tag to
> the protein which facilitates purification over an affinity column. I
> have used numerous his tag fusion vectors but am looking to try
> something new. Please respond by e-mail.
Have you tried GST-fusion vectors? GST is a protein which binds with high
affinity to Glutathione-S-Transferase, which you can purchase bound to
beads fro bead purification. To elute off you compete with glutathione,
which can be dialyzed out of your elution buffer.
There is a line of these vectors available from Pharmacia (and others sell
them too). Look for the pGEX line of vectors. They vary in the usual
ways (selection marker, etc...) and also in the length of the linker region
between the GST and the polylinker. Some have a thrombin site engineered
inbetween GST and the polylinker so you can cleave the GST off after
They work like a charm.
More information about the Methods