yeast strain for disr

Tod Lorenz tod.lorenz at ghawk.com
Tue Jan 31 20:59:00 EST 1995


 -=> Quoting Colin Macdiarmid to All <=-

 CM> Hi out there
 CM> I need some help from experienced gene-disruptors.  I have two genes
 CM> which I need to interupt, and want to do this by insertion of the URA3
 CM> gene.  After that I want to be able to use 5-FOA to reselect for a
 CM> strain which has excised the URA3 gene (but retained my disrupted
 CM> version of the gene).  The genes were both from a library made from the
 CM> S288C strain, and strains with this background are sensitive to low
 CM> concentrations of 5-FOA on plates with proline as sole nitrogen source.
 CM> So ideally I want a diploid yeast strain derived from s288C to do my
 CM> disruptions.  Anyone have an idea which strain to use? Can I get it
 CM> from the YGSC? Please post replies or send direct to me.
       Is the gene(s) disruption lethal?  It should not matter what
  the source of the clone bank is, just the background of the yeast
  being transformed...try the Berkeley yeast stocks for other ura3
  strains to use.  Why the need for proline as a nitrogen source
  out of curiosity?  Why not just excise the URA3 gene out with a
  restriction enzyme if the plasmid is mapped and get around all
  the FOA problems?

	  /^\/^^^^\/^\     R. Todd Lorenz
	    | #  @ |      Ellicott City, MD
	     \ :: /      T.LORENZ at GENIE.GEIS.COM
	     =\||/=
	       #@



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