Hi,
We are trying to clone the gene which complements a mutant, but have
failed to clone it out of a CEN library. We know that the gene is CEN
linked, and we are entertaining the possibility that we can not clone the
gene because the gene would bring a CEN, and two CENs is toxic for a
plasmid.
First question: Do others have experience with this problem (and any
solutions)?
We could map the gene by crossing to strains with CEN linked markers, but
this will prove difficult to analyze because a reporter present in our
strain (and absent from the mapping strains) is needed to determine the
phenotype.
We have another idea: if we can get disruptors (for non essential genes)
for each of the chromosomes, we can easily make strains to do the mapping
in a strain containing our reporter.
If you have a disruptor (URA3 preferred) for a non essential CEN linked
gene, please contact me.
We know our gene is not on chromosome III (LEU2) or IV (TRP1).
Thanks.
David Stillman
Department of Cellular, Viral and Molecular Biology
University of Utah Medical Center
Salt Lake City, UT 84132
stillman at bioscience.utah.edu
phone: 801-581-5429
fax: 801-581-3607
