problems with GST-fusion protein

R. John Lye rjl6n at virginia.edu
Fri Sep 1 08:47:07 EST 2000


Silke Meister wrote:

> I cloned my protein in the pGEX-vector and wanted to purify this
> GST-fusion protein, but I wasn't succesful!
> The protein didn't bind to the sepharose-beads. In my opinion the
> protein prevent the binding from GST to GSH, because the empty
> GST-vector without the protein can bind!

First, have you checked that the fusion protein is soluble rather
than in inclusion bodies?  If it is in the pellet, you're unlikely to
get much binding to the beads.

John








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