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2 VERY SIMPLE yeast questions

bio1 at mail.com bio1 at mail.com
Mon Jan 24 10:11:10 EST 2000


Hello All,

Maybe someone could help me... I am using the S. Cerevisiae complementation 
system to screen for the proteins that complement the deletion of the 
protein "X" in my strain of yeast . What I want to do is to knockout 
another gene in this strain (non-essential under normal conditions) to see 
if some of the proteins that normally rescue would require this gene to 
work.  The gene "X" is disrupted by the insertion of HIS marker, and cells 
are maintained alive by the URA plasmid expressing the protein "X". I 
introduce the new genes on TRP plasmid and then select on 5-FOA. So what 
would be the best way to do it and the best marker to use?

Question 2 is about the same experimental system. I am interested in seeing 
if the introduction of the new gene "Y" instead of "X" causes the Unfolded 
protein response induction in the cells. Are there any plasmids that allow 
you to introduce some reporter under the control of the UPR promoter and 
see if the UPR is being induced?

thanks in advance for any help,

Anton.

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