Pichia expression

[Kathryn Graham]

	I am one of two students in a lab trying out the Pichia Pastoris
expression system from Invitrogen.  We have been trying with three
different proteins, in two different vectors (pHIL-D2 and pHIL-S1).  So far
we are not seeing any protein production.  I have screened 32 colonies
which were His+, and looked like the AOX was gone, but nothing so far.  I
am currently isolating DNA so that I can test by PCR to see if my cDNA has
integrated at the AOX locus.  At this point I can't say why we aren't
getting any protein, but it seems that the procedure is a little more
complicated than Invitrogen advertises.  If anyone out there has any
suggestions for me, let me know.

							Thanks			
							Piers Nash

mcf-p at bones.biochem.ualberta.ca