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Leu2-d and Ura3-d

Wed Nov 9 10:27:11 EST 1994

Dear All

Has anybody used the Leu2-d or Ura3-d markers to get multicopy 
expression ?
We have transformed these into strains that are very competent for 
transformation of other muticopy and CEN vectors, as well as for 
integrative transformations.
We find that Leu2-d does not transform directly onto -leu media, even 
if left for a week as suggested in the originators paper. However, if 
another marker is present on the plasmid ( URA3 or TRP1 ), then we can 
transform onto this media and get sub-clones to grow on -leu. This 
sort of defeats the purpose that we want it for.
The Ura3-d transforms directly onto -ura, but as the reference 
provided by ATCC does not give information on copy number, I was 
wondering if anyone had an idea, before we go ahead and check.

So, any previous experience with these ?

      Dave Pearce  DPEA at bphvax.biophysics.rochester.edu

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