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spore germination on YEP-galactose

John Cannon cannon at MEDSCI.MBP.MISSOURI.EDU
Fri Mar 4 20:28:17 EST 1994


The question refered to dissecting tetrads on YEP-galactose to allow
spores with presumably a gene-disruption to grow in the presence of
a gene under galactose control. We have used this technique and have found
no problems for spores germinating on galactose, mannose, fructose,
and a variety of nonfermentable carbon sources. Your spore inviability
is potentially due to your particular gene / strain combination and not
a general finding. Two things I can think of are to check whether your
mitotically growing cellsdiploid transformants suffer any viability loss
when you induce with galactose. In particular, is the plasmid retention
the same on galactose as it is on glucose? This could indicate that
hyper-expression is detrimental to viability. Second, it might make a
difference whether you are using _gal2_ strains. Mitotically growing
_gal2_ strains can grow on 2% galactose plates OK, but this might
be problem in germinating spores. A lot of lab strains are _gal2_ !

John Cannon
Department of Molecular Microbiology and Immunology
University of MIssouri
Columbia, MIssouri



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