YPD medium lacking uracil

John McCusker mccusker at ABACUS.MC.DUKE.EDU
Mon Apr 21 10:00:51 EST 1997


I don't know of any YPD like medium that lacks uracil but there may be a
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).  I've never tried this technique but it is
published

Loison, G., M. Nguyen-Juilleret, S. Alouani and M. Marquet. 1986.
Plasmid-transformed ura3 fur1 mutants of S. cerevisiae: an autoselection
system applicable to the production of foreign proteins.  Biotechnology 4:
433-437.

Good luck

*****************************************************************************
John H. McCusker, Asst. Prof.
Dept. of Microbiology, 3020
Duke University Medical Center
Durham, NC 27710

phone:  (919) 681-6744
fax:    (919) 684-8735
e-mail: mccusker at abacus.mc.duke.edu





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