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In search of a delta-trp1 plasmid

Adam Gower agower at biochem.umass.edu
Tue Nov 11 11:21:00 EST 1997


I am in search of an integrative plasmid, carrying a delta-trp1
deletion, with a URA3 marker, to be used for a two-step deletion of TRP1
in Sc252 derivatives.  We have been using plasmid pNKY1009, which
carries a TRP1::hisG-URA3-hisG (Alani, Cao, and Kleckner 1987, Genetics
116:514-545) construct that can be cured of URA3 by recombination
between the hisG cassettes, leaving a single hisG as a disruption of
TRP1.  However, the yield we have obtained with this process in these
strains has been extremely poor.
	If anyone has a plasmid of this kind, I would greatly appreciate a
reply.

										Thanks,

												Adam Gower






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