Sat May 15 14:49:00 EST 1993

Hi Tom!
    Why don't you just knock out the HIS3 gene using
the one step deletion method of Rothstein (see Meth. Enzym. Vol 194 pp;281)
I have a URA3 deletion plasmid you can use if you like.
then you know that there is no HIS3 activity. If you use 
the highly efficient transformation method of Gietz et al. (1992)
N.A.R. 20:pp 1425. you should have no problems getting this deletion
to go!  Drop me a line if you need this plasmid.
Dan Gietz
R.Daniel Gietz Ph.D.
Assistant Professor
Department of Human Genetics
University of Manitoba
770 Bannatyne Ave, Rm 250
Winnipeg, Manitoba, Canada
R3E 0W3
Tel.: (204)788-6458
Fax.: (204)786-8712

REPLY FROM: Gietz, Dr. Dan
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From: larsont at
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Date: Fri, 14 May 93 18:00:08 EDT
To: yeast at
Apparently-To: yeast at
Subject: aminotriazole
We're using 3-amino-1,2,4-triazole (ATA) to inhibit activity of the HIS3
gene product and cause auxotrophy on minimal medium.  Our yeast seems to
be resistant to our ATA.  Any suggestions?  The ATA is from Sigma.

Thom Larson

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