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YPD medium lacking uracil

Vershon, Andrew K vershon at OCELOT.RUTGERS.EDU
Tue Apr 22 10:14:50 EST 1997

John McCusker writes: 
<A fur1 strain that contains a chromosomal ura3 mutation *and* URA3 on a plasmid
is <absolutely dependent for growth on the URA3 plasmid -- it dies if it loses
<plasmid.  This has nothing to do with 5FOA.

In doing a synthetic lethal screen using only URA3 as the marker on the plasmid
with our gene of interest, we picked up (as an artifact) a mutant in FUR1.  This
mutant failed to grow if we lost the URA3 on the plasmid - even in the presence
of uracil.  However a fur1 ura3 strain can grow if supplemented with uridine
instead uracil in the media.  The cell uses the URK1 gene product to convert
uridine to uridine-5'-phosphate, the downstream product of both URA3 and FUR1.  
 (We also picked up CAN1 as a suppressor in this screen- but that is another
artifact disaster). 

Drew Vershon
Waksman Institute
Rutgers University
(908) 445-2905
vershon at mbcl.rutgers.edu

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