25 Jun You have been hired by the bakery industry to find a transporter
Question
1. You have been hired by the bakery industry to find a transporter gene that can confer yeast cells resistance to a toxin. You have access to a huge, unexplored years cDNA library. How would you find and clone the gene that codes for the toxin transporter? Be sure to include controls.
2. The gene wha is normally transcribed in humans in response to serum testosterone. A man shows no expression of gene wha protein, even though he makes normal levels of testosterone in his blood. Propose two different possible mutations that could independently produce this lack of androgen response by the wha gene. For each proposed mutation, detail what locus or DNA site is mutated and how the mutation would produce the described phenotype.
3. In a certain human cell line an enzyme gene that is normally expressed in human cells is not present. We know this because we have tested the cell culture for activity of the particular enzyme. List 4 different hypotheses for why the enzyme is not being made in the cell line, and briefly tell what experimental method(s) you might use to test each hypothesis.
4. With restriction enzyme “mapping”, you create abundant samples of the following short fragments of the full DNA:
ACCTAGATCATGC
TGGCAATAGCTAATA
GATACCTA
CATGCCGACATGGC
AATATAG
Use gene walking analysis to reconstruct the original DNA strand.
b. Examine the strand you figured out above and state wether this sequence looks most like part of a protein-coding sequence or promoter sequence.
c. What clues from the strand of DNA above suggested the answer to b?
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