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Cycloheximide resistance

Noelle Yans s931905 at student.ulg.ac.be
Thu May 18 14:36:34 EST 2000


Hi yeast people,

I'm trying to make a genetic screening system similar to the reverse two-hybrid system. Therefore, I use a plasmid expressing the cyh2 wild type allele under the control of the GAL1 promoter. This construct has to be stably integrated at the leu2 locus. Before that, I have to make my yeast strain resistant to cycloheximide (my strain is W303-1A, haploid, with five auxotrophic markers : leu2 ura3 his3 trp1 and ade2). As I am quite new to yeast work, can anyone tell me what are the best conditions :
    1) to select for yeast cells resistant to CHX (which CHX concentration to use for the selection and after....)
    2) to ensure that the correct phenotype will be maintained throughout further manipulations

Thank you in advance

NoelleYANS
PhD student
University of Liège
Laboratory of Molecular Biology and Genetic Engineering
Alle du 6 Aout, 17
4000 Liège
BELGIUM




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