expression/fusion vectors

[Anton Tutter]

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 
purification.

They work like a charm.


Good luck,

Anton Tutter
Salk Institute