foreign protein expression in yeast

lledgerwood at chmeds.ac.nz lledgerwood at chmeds.ac.nz
Wed Jan 12 21:09:13 EST 1994


Hi 
I'm working on a project expressing preproalbumin and various mutants. I'm
using rwo strains - AB110 and AB118, which are both ura3-52 and AB118 is
Kex2-. My problem is very low expression levels. The vector is pAB23
which has URA3 selection and expression is under the control of a GAP 
promoter and terminator.  I am presently growing the yeast in 0.67% 
yeast nitrogen base without amino acids, 0.5% casamino acids and 2% 
glucose at 29C, 350rpm overnight (although I have tried growing up to 48h.

I am expecting the albumin to be secreted into the medium, and I have tried
extracting the yeast pellet (with glass beads) - I got protein out but no 
albumin in significant amounts.

Does anyone have any advice as to what yields I should expect and where 
I am going wrong. 

Liz Ledgerwood

lledgerwood at chmeds.ac.nz

Christchurch School of Medicine
Christchurch
New Zealand




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