Pichia recombinant proteins

BSpinoza bspinoza at aol.com
Thu Jul 6 20:02:17 EST 1995


         I have been studying the human erythropoietin receptor
extracellular domain as a bacterially expressed GST fusion protein. I
became unsatisfied with the activity of this recombinant protein- only 1%
of the soluble protein bound ligand with the expected high affinity, even
after tedious affinity purification.   Consequently, we have expressed
this protein in the yeast Pichia pastoris.  It seems to have a much higher
specific activity than the bacterially expressed protein (still working on
how much...) and is appropriately glycosylated (by molecular weight). 
Unfortunately the yields are, to say the least, disappointing.  
           I have three questions:
1.  Are a lot of people using this system (Invitrogen)?
2.  What is the experience with improving yields?
3.  Anyone have any ideas for an interesting collaboration with this
receptor fragment?

Kevin Harris- harrisk at uthscsa.edu 



More information about the Proteins mailing list