You are part of a yeast genetics research team studying on the regulatory mechan

Question

You are part of a yeast genetics research team studying on the regulatory mechanism of polarized growth. When you expressed yeast genomic DNA library (in plasmid) in the yeast mutant strain showing polarized growth defect, some strains reverted back to wild type polarized growth. To identify the genes play important role in governing the polarized growth , you will need to isolate the plasmid holding genomic DNA fragment from the strain with reverted polarized growth, and analyze the segment of DNA in the plasmid.

How would you perform this research? (Describe the experimental and bioinformatics procedures to find out the gene information related to the phenotypic reversion).

Explanation / Answer

Isolate the plasmids from the reverted mutants and restriction digest the plasmid to release the genomic DNA fragment.

Sequence this genomic DNA fragment to identify the changes happened due to mutations.

Run comparative analysis between the genomic DNA isolated from reverted mutants and the original genomic DNA inserted to observe the changes that gave rise to these reverted mutants.

Then from the comparative analysis, we could say whether this was a point or frameshift mutation which gave a phenotypic revertant mutant.

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