timed induction of gal1 promoter
Soon A. Lee
s.lee at auckland.ac.nz
Thu Feb 29 15:44:37 EST 1996
Dear net readers,
I wonder if you could help me. My problem is this - I want to do northerns/growth curves on strains overexpressing certain genes from the gal1 promoter. The overexpression decreases the growth rate, meaning I can't harvest the strains all at once at log phase for analysis (if I grow from scatch on galactose). I would like to induce the promoter at say OD600 0.3-0.4, then allow four hours more growth for induction before harvest. What is the best way to do this? I have heard of growing on glycerol/lactate medium followed by galactose addition at the appropriate time (anyone have a description of glycerol/lactate synthetic medium?). However, for certain reasons I would prefer to use a fermentable substrate. Could I make medium with a low glucose conc, with normal galactose, and rely on depletion of the glucose for induction? How much glucose should I use to see depletion by about early log phase? Any tips would be much appreciated.
University of Auckland
More information about the Yeast