mallory lutz wrote:
>> Hi. I have a yeast gene that is essential for life on glucose. I'm
> planning to use a sectoring assay as my method of choice for screening
> for PCR generated ts mutants. My gene is already deleted in S.
> cerevisiae strain W3031A. This would work fine for everything except it
> is ADE3. Does anyone have a W3031A or W303 strain that is ade3? My
> strain is mating type a, so I could use a strain mating type alpha.
> Thanks, Mallory
Sure. I've made a W303 strain that has a deletion of almost all of
ADE3. Both mating types. Called YMW1 and YMW2.
See Zieler, Walberg and Berg 1995. MCB15, 3227.
I've also got some ADE3 plasmids if you need them. The ADE3 gene on the
plasmids does not span the region deleted, so gene converting the ADE3
deletion back to wild type ADE3 doesn't give false positives.
Mark Walberg walberg at simmons.swmed.edu