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YPD medium lacking uracil

Tritech Research Tritech_Research at lamg.com
Tue Apr 22 08:00:01 EST 1997

John McCusker said:
> simple way for you to demand retention of your URA3 plasmid.  Try taking
> your transformed (Ura+) strain and selecting for a 5-fluorouracil resistant
> mutant.  The result will be a strain that is deficient in the pyrimidine
> salvage pathway (5FU resistant) and is completely dependent for growth on
> the de novo pyrimidine synthesis pathway (which means that anything that
> loses the URA3 plasmid dies).

stahl at infobiogen.fr says:

Perhaps I didn't understand the way you explained this, so I will transcribe
reality in an easier way :

Everything which is [Ura+] is [5FOA sensitive]
Everything which is [5FOA resistant] is [Ura-] (in fact ura3- or ura5-)

So I think this is the contrary of what Colin MacDiarmid asked for... (e.g.
selection in rich media). Anyway, I don't remember the reason, but 5FOA
selection is 
always done on minimal media.
What you say is right for 5-fluoroorotic acid (5FOA), but 
5FOA is not the same as 5FU, so the idea might work.

Andy Papp
tritech_research at LAMG.COM

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