Difference between revisions of "Cdomin12 Week 6"

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==Acknowledgments==
 
==Acknowledgments==
 
1. I worked with
 
1. I worked with
 +
 
2."Except for what is noted above, this individual journal entry was completed by me and not copied from another source."
 
2."Except for what is noted above, this individual journal entry was completed by me and not copied from another source."
  

Revision as of 16:00, 6 October 2019

Contents

Purpose

Questions

1. (Question 5, p. 110) Choose two genes from Figure 4.6b (PDF of figures on Brightspace) and draw a graph to represent the change in transcription over time. You can either create your plot in Excel and put the image up on your wiki page or you can do it by hand and upload a picture or scan.

2. (Question 6b, p. 110) Look at Figure 4.7, which depicts the loss of oxygen over time and the transcriptional response of three genes. These data are the ratios of transcription for genes X, Y, and Z during the depletion of oxygen. Using the color scale from Figure 4.6, determine the color for each ratio in Figure 4.7b. (Use the nomenclature "bright green", "medium green", "dim green", "black", "dim red", "medium red", or "bright red" for your answers.)

Gene X

1 hour:black

3 hours:dim red

5 hours:black

9 hours:dim green

Gene Y

1 hour:black

3 hours:dim red

5 hours:medium green

9 hours:bright green

Gene Z

1 hour:black

3 hours:dim red

5 hours:dim red

9 hours:dim red

3. (Question 7, p. 110) Were any of the genes in Figure 4.7b transcribed similarly? If so, which ones were transcribed similarly to which ones?

Gene X and Y were transcribed similarly in increase in oxygen and than loss in oxygen.

4. (Question 9, p. 118) Why would most spots be yellow at the first time point? I.e., what is the technical reason that spots show up as yellow - where does the yellow color come from? And, what would be the biological reason that the experiment resulted in most spots being yellow?

Most spots would be yellow at the first time point due to the equal ratio of green:red or repression:induction. No transcription has yet to occur so it does not appear as either red or green but yellow to indicate both are present in equal ratio.

5. (Question 10, p. 118) Go to the Saccharomyces Genome Database and search for the gene TEF4; you will see it is involved in translation. Look at the time point labeled OD 3.7 in Figure 4.12, and find the TEF4 spot. Over the course of this experiment, was TEF4 induced or repressed? Hypothesize why TEF4’s change in expression was part of the cell’s response to a reduction in available glucose (i.e., the only available food).

TEF4 was repressed over the course of the experiment. This change in expression was due to its role in translation and the binding of tRNA to the ribosome. As less glucose becomes available, the cell cannot make more energy and thus it stops the cellular expression of certain genes, including TEF4. There is not enough energy to continue with translation and the production of proteins.

6. (Question, 11, p. 120) Why would TCA cycle genes be induced if the glucose supply is running out?

TCA cycle genes would be induced due to

7. (Question 12, p. 120) What mechanism could the genome use to ensure genes for enzymes in a common pathway are induced or repressed simultaneously?

8. (Question 13, p. 121) Consider a microarray experiment where cells deleted for the repressor TUP1 were subjected to the same experiment of a timecourse of glucose depletion where cells at t0 (plenty of glucose available) are labeled green and cells at later timepoints (glucose depleted) are labeled red. What color would you expect the spots that represented glucose-repressed genes to be in the later time points of this experiment?

9. (Question 14, p. 121) Consider a microarray experiment where cells that overexpress the transcription factor Yap1p were subjected to the same experiment of a timecourse of glucose depletion where cells at t0 (plenty of glucose available) are labeled green and cells at later timepoints (glucose depleted) are labeled red. What color would you expect the spots that represented Yap1p target genes to be in the later time points of this experiment?

10. (Question 16, p. 121) Using the microarray data, how could you verify that you had truly deleted TUP1 or overexpressed YAP1 in the experiments described in questions 8 and 9?

Acknowledgments

1. I worked with

2."Except for what is noted above, this individual journal entry was completed by me and not copied from another source."

Cdomin12 (talk) 16:57, 6 October 2019 (PDT)

References

  • Campbell, A.M. and Heyer, L.J. (2003), “Chapter 4: Basic Research with DNA Microarrays”, in Discovering Genomics, Proteomics, and Bioinformatics, Cold Spring Harbor Laboratory Press, pp. 107-124.