You are studying the Drosophila Tral gene, whose protein product is involved in mRNA localization. You want to examine the regulation of Tral expression in flies. 1. To facilitate your study, you create a GFP tagged version of Tral. Which one of the following is not a step you would perform to accomplish this?
A. PCR the Tral gene from Drosophila genomic DNA.
B. Digest the PCR product and a GFP-containing plasmid.
C. Add the insert and vector together and add DNA ligase.
D. Transform the ligation mixture into Drosophila.
E. All of the above would be performed.
2. For this cloning, we need to include a promoter in the plasmid:.
A. Bacteria.
B. Drosophila.
C. Neither.
D. Both.
3. You also speculate that various transcription factors could be regulating Tral expression One such factor is Snail.
a. If Snail is a transcription factor activating Tral expression, which of the following could potentially be a function of Snail?
A. Inhibit the binding of RNA polymerase at the Tral promoter.
B. Recruit a histone modifying protein that opens up the chromatin at the Tral gene.
C. Recruit DNA helicase at the origin of replication.
b. You want to determine whether Snail binds to the Tral promoter DNA. Which technique(s) would allow you to determine whether this occurs.
A. Quantitative RT PCR.
B. Chromatin Immunoprecipitation.
C. Immunoprecipitation.
D. Western blot.
E. DNA footprinting.
4. Now you want to determine the effect of the loss of Snail on Tral-GFP expression. If Snail is an activator of transcription, draw/describe the expected results from a:.
A. Northern blot.
B. Q-RT PCR reaction.