[Yeast] EMM + glycerol?

Nathan Blewett via yeast%40net.bio.net (by blewettn At mail.nih.gov)
Tue Jan 30 13:02:51 EST 2007


Hello,

I have deleted a gene, Maf1, in 2 S. pombe strains, this results in a growth
defect at 37C on YES 3% glycerol/ no glucose.  I would like to transform
maf1 back into the deleted strain on an expression vector to rescue this
phenotype, as well as the anti-suppression phenotype.  Regarding the growth
defect, I've constructed the vector, which is pREP4X expressing maf1 under a
thiamine repressible promoter, and confirmed that I am making the protein
via western.  However, when I plate on EMM w/o glucose, with glycerol 3% ,
there is no growth, even from the parent strain yYH1, which contains the
ade6-704 allele and a suppressor tRNA, but is quite happy otherwise.

Does anyone know why pombe would be unable to grow on EMM containing a
non-fermentable carbon source?  Essentially, I need to plate on media
lacking thiamine, or put maf1 under its' own promoter, but I am curious
about the inability to grow on EMM glycerol.  Thank you.

Nathan Blewett



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