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plasmid to make trp1 strain

jansma at RESUNIX.RI.SICKKIDS.ON.CA jansma at RESUNIX.RI.SICKKIDS.ON.CA
Mon Feb 14 22:24:22 EST 1994


Hi yeasters,
        I wonder if somebody out there might be able to help me out.  I
have a 
strain that is prototrophic for tryptophan and I would like to introduce
a trp1 mutation into it.  Normally, I would just cross it with a trp1
strain
and screen for trp- in the spores.  In this case though, I want to be
particularly careful to maintain the original background.  Also, the strain
has three different genes deleted in the chromosome with no markers to 
follow them, making the tetrad analysis more difficult.  Does anybody out 
there have a plasmid that introduces a trp1 allele (preferably one with a
deletion) into the TRP1 locus by a two-step approach?  More specifically,
an 
integration plasmid with a delta-trp1 allele that has a suitable unique
restriction enzyme site in it, such that the plasmid could be directed to
the TRP1 locus as well as a URA3 marker such that only the trp1 allele 
might be left behind after treatment on 5-FOA?
Thanks for your attention,

David Jansma
jansma at sickkids.on.ca

David Jansma
c/o J.D. Friesen
Hospital for Sick Children
555 University Ave.
Toronto, Ontario
M5G 1X8
Canada 




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