Any hints on recombinant protein expression in Pichia?

Dima Klenchin klenchin at facstaff.REMOVE_TO_REPLY.wisc.edu
Mon Nov 15 14:21:41 EST 1999


We are thinking about trying the system out for the mass production of
a 150 KDa cytosolic protein. Unfortunately, I don't know enough about it 
to be able to judge manufacturer's claims about it. I would greatly 
appreciate any hints, comments or suggestions related to the use of
this system. I searched DejaNews and there are only few hits there. 
Among many questions I don't have clear answer to are:

Does anyone besides Invitrogen make the kits/expression vectors?
What vectors (pPICZ, pGAPZ, etc) may work better? 
Is proteolysis indeed a very serious problem with yeast expression?
What kind of advantage vectors for secretable epression offer? Is 
proteolysis a bigger probelm or a lesser one with them? 
What could be the "real life" yelds in case of ~ 150 kDa proteins? 

Thanks! 
        Dima





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