1. You have identified a yeast mutant in gene1 that results in a it does not gro
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Question
1. You have identified a yeast mutant in gene1 that results in a it does not grow at 37 C. You know the identity and sequence of gene 1, but you do function of the gene. temperature sensitive phenotype, i.e enc a. Describe the specific steps of an experiment that you would carry out to identify suppressors of the mutant You successfully identified a suppressor (mutated in gene 2) that has a very strong phenotype, i.e., it completely suppresses temperature sensitivity, such that the double mutant (with mutations in gene 1 and gene 2) grows very well at 37°C. You would now like to identify gene 2. How will you do this?Explanation / Answer
a. Generate and isolate single mutants (say by using EMS as the mutagen) --> along each mutant to mate with gene1 mutant --> observe the growth of each of these double mutants formed at 37°C. The one which survives has a suppressor mutation. (Note sequence gene1 to ensure that the suppress mutation is not in gene 1 itself)
b. Take the genomic library of yeast and transfect the double mutant with all clones in the library
Rescue (temperature sensitivity) obtained of whichever clone would correspond to gene 2 / suppressor
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