Dear friend,
I am trying to use yeast strain BJ2168 to over-express Gal1 promoter driven GFP. I grow the strain in SC complete media-glucose to OD 0.5. After washing the cell with PBS, I replace the media with SC-galactose. The cell seems stop growing after that. My question is that how long the cell would recover from the carbon resource swopping, normally in your hand?
thank you.
ziguo
Hi there,
To avoid this problem of lag phase, you should grow cells in raffinose containing media (the rate is slower than with glucose). The simple addition of galactose will trigger GFP expression instantly without affecting the growth rate.
Ziguo,
To build on Dominque's suggestion, dextrose represses GAL1 expression, so using a non-inducing sugar such as raffinose to grow the yeast in prior to galactose induction will also eliminate problems with poor initial induction after shifting to galactose.
Certain strains do have a difficult time proliferating in raffinose medium, however, so test yours first.
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