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
