Tetrahymena thermophila is a great organism to study DNA damage repair because it has a great DNA repair machinery once it is able to reconstruct its genome during replication (it is completely degraded). The isolated gene, Wd40, was the studied gene that was determined by bioinformatics programs and we wished that this isolated gene was related to the DNA repair machinery, but it unfortunately wasn’t. We didn’t actually pick this gene.
Hi Shriya, good questions. The things that you asked were actually the two choices out of our hands. T. thermophila was the model organism for our entire lab, we did not choose it. The organisms we used were cultured and grown by our TAs. The Wd40 protein was assigned to our group at random from a database of previously unstudied T. thermophila genes.
You mention in your conclusion that your loading controls were off, effecting your bioinformatic quantitative analysis. If you had more time and were able to correct for this, how do you believe your results may change?
Even though the loading controls were off, the answer to our initial question would not change: Wd40 gene is not part of the Tetrahymena thermophila DNA repair machinery. The expression did not increase after DNA damage. Some further analysis can still be made, so it is of our interest to have the most accurate data as possible, once the actual function of Wd40 still remains unknown. Nonetheless, it would not change our results.
It is probably because Wd40 is not part of the DNA damage repair machinery. If it was, it’s expression would increase after DNA damage. Nonetheless, we can still use the results from this experiment to explore Wd40 function.
Hi Sophia. I believe that we did not see an increase in expression as Wd40 is likely not involved in the repair pathway. While we didn’t see upregulation, we also did not see downregulation. This is important because it points to the conclusion that Wd40 is completely uninvolved -its expression is completely detached from DNA damage.
Why did you guys pair Wd40 with the DNA damage repair pathway in the first place? Why did you guys think that they would have a connection with eachother?
Good question Ian. The gene Wd40 itself is part of DNA damage repair and after using bioinformatic programs, the gene isolated from Tetrahymena thermophila was Wd40. The purpose of this lab is to find new genes that help with this organism DNA damage repair and we hoped it had. However, as it was seen, the Wd40 gene expression did not increase after DNA damage.
curesymposium.org 
Great presentation! I was curious on where you guys sourced Tetrahymena thermophila and why you picked to match it with the Wd40 protein? Thank you!
LikeLike
Tetrahymena thermophila is a great organism to study DNA damage repair because it has a great DNA repair machinery once it is able to reconstruct its genome during replication (it is completely degraded). The isolated gene, Wd40, was the studied gene that was determined by bioinformatics programs and we wished that this isolated gene was related to the DNA repair machinery, but it unfortunately wasn’t. We didn’t actually pick this gene.
LikeLike
Hi Shriya, good questions. The things that you asked were actually the two choices out of our hands. T. thermophila was the model organism for our entire lab, we did not choose it. The organisms we used were cultured and grown by our TAs. The Wd40 protein was assigned to our group at random from a database of previously unstudied T. thermophila genes.
LikeLike
You mention in your conclusion that your loading controls were off, effecting your bioinformatic quantitative analysis. If you had more time and were able to correct for this, how do you believe your results may change?
LikeLike
Even though the loading controls were off, the answer to our initial question would not change: Wd40 gene is not part of the Tetrahymena thermophila DNA repair machinery. The expression did not increase after DNA damage. Some further analysis can still be made, so it is of our interest to have the most accurate data as possible, once the actual function of Wd40 still remains unknown. Nonetheless, it would not change our results.
LikeLike
Great Presentation! Why do you think expression did not increase after DNA damage with the PCR test.
LikeLike
It is probably because Wd40 is not part of the DNA damage repair machinery. If it was, it’s expression would increase after DNA damage. Nonetheless, we can still use the results from this experiment to explore Wd40 function.
LikeLike
Hi Sophia. I believe that we did not see an increase in expression as Wd40 is likely not involved in the repair pathway. While we didn’t see upregulation, we also did not see downregulation. This is important because it points to the conclusion that Wd40 is completely uninvolved -its expression is completely detached from DNA damage.
LikeLike
Why did you guys pair Wd40 with the DNA damage repair pathway in the first place? Why did you guys think that they would have a connection with eachother?
LikeLike
Good question Ian. The gene Wd40 itself is part of DNA damage repair and after using bioinformatic programs, the gene isolated from Tetrahymena thermophila was Wd40. The purpose of this lab is to find new genes that help with this organism DNA damage repair and we hoped it had. However, as it was seen, the Wd40 gene expression did not increase after DNA damage.
LikeLike