While conducting a genetic screen to identify new developmental genes, you have
ID: 190213 • Letter: W
Question
While conducting a genetic screen to identify new developmental genes, you have found an embryo that has severe posterior defects. You call this gene rumpless, which was found to encode a transcription factor protein. The professor you are working for gives you an exciting project to study the rumpless gene for your undergraduate summer research project. a. Describe an experiment you could perform to determine when and where rumpless is expressed. b. You have identified a 500-nucleotide piece of DNA that might be an enhancer sequence for rumpless gene expression in the posterior region of the embryo. Describe how you could test whether this enhancer is necessary and sufficient for rumpless transcription. Describe both the experiments and the results that would support your hypothesis that the enhancer is both necessary and sufficient.Explanation / Answer
a.
To find out the temporal and spatial expression domain of rumpless, we can fuse it to a reporter gene such as GFP and observe its expression.
b.
Make a mutant in which enhancer is remover and observe for the expression of the gene (Tagged to a reporter).
If enhancer is sufficient for the gene expression, we would find reduced/no GFP expression in the mutant.
Take a loss of function mutation at the rumpless locus (in which the mutation is in the coding sequence), try to complement it with the mutant version (in which enhancer is deleted).
If complementation does not occur, it shows that enhancer is necessary for the gene expression.
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