If I want to screen for mutant yeast (S. cerevisiae) colonies that have defects in mitotic chromosome condensation, can I screen for colonies arrested in Mid-M phase? Would that work?

  • $\begingroup$ It is going to need to be a temperature sensitive mutation, so that you can get them to grow at the permissive temperature and arrest during mitosis at the restrictive temperature, because otherwise your cells would not grow. $\endgroup$ – AMR Oct 18 '15 at 4:43
  • $\begingroup$ You probably would also want to do a thorough research review of Leland Hartwell and Paul Nurse's work as they worked out the cell cycle using yeast. $\endgroup$ – AMR Oct 18 '15 at 4:47
  • $\begingroup$ The ts mutation would be like a "primary screen" from there one can do a more sensitive secondary screen? Thank you! $\endgroup$ – player87 Oct 18 '15 at 15:03
  • $\begingroup$ Right. So you find the ts mutant, and then you isolate mRNA from population grown at permissive temp and population grown at restrictive temp. Then you make cDNA and run that out on a microarray (They probably have plenty of whole genome chips available for yeast). You can then see the genes that are down (or up) regulated from a comparison of the two arrays, and that should tell you where gene expression has changed. You will then need to make probes for those genes and do FISH to try and determine where they are localized in the nucleus. $\endgroup$ – AMR Oct 18 '15 at 15:54

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