Hypr2c was expected to have a brighter band in the +HU (treated) lane as I expected it to be a part of the DDR pathway therefore meaning it would have higher expression! However, I got no presence of bands 🙁
These unexpected results just mean there was no expression of Hypr2c after DNA Damage was induced! This was just unexpected because I hypothesized there would be more expression due to its similarity to Rad51 in the gene expression profile and therefore have a role in DDR pathways, but there were no bands, just meaning it is not a part of DNA Damage repair.
In the future, I would hope to use protein localization experiments where I could actually tag Hypr2c in a DNA sequence and use this tag to see where it localizes within the cell. From this, I could then use its location to run more experiments like the ones used in this lab, to determine its function!
Ftt18 was our positive control so it doesn’t necessarily mean that it is involved in DNA Damage repair but it was used because it is known to anneal in every Tetrahymena gene, so it was used as a basis of showing if the experiment was valid. If done properly, Ftt18 will always show bands when using Tetrahymena genes.
As of right now, I’m not sure! When researching Hypr2c, there was no known information about it (no known connections to other proteins) so right now all we know is that it is a hypothetical protein with similar peaks to Rad51. We now also know that it is not involved in DNA Damage repair so we can search for other possible functions!
What where the expected results for Hypr2c in figure 2?
Hypr2c was expected to have a brighter band in the +HU (treated) lane as I expected it to be a part of the DDR pathway therefore meaning it would have higher expression! However, I got no presence of bands 🙁
I may have missed this but do you have any ideas as to why you got unexpected results?
These unexpected results just mean there was no expression of Hypr2c after DNA Damage was induced! This was just unexpected because I hypothesized there would be more expression due to its similarity to Rad51 in the gene expression profile and therefore have a role in DDR pathways, but there were no bands, just meaning it is not a part of DNA Damage repair.
How do you plan on testing Hypr2c in different ares of the cell if given the chance to continue on with testing?
In the future, I would hope to use protein localization experiments where I could actually tag Hypr2c in a DNA sequence and use this tag to see where it localizes within the cell. From this, I could then use its location to run more experiments like the ones used in this lab, to determine its function!
Since you concluded that hyper2c was not expressed for DNA repair, does that mean that fit18 is because it showed positive results?
Ftt18 was our positive control so it doesn’t necessarily mean that it is involved in DNA Damage repair but it was used because it is known to anneal in every Tetrahymena gene, so it was used as a basis of showing if the experiment was valid. If done properly, Ftt18 will always show bands when using Tetrahymena genes.
What other functions do you think your gene has?
As of right now, I’m not sure! When researching Hypr2c, there was no known information about it (no known connections to other proteins) so right now all we know is that it is a hypothetical protein with similar peaks to Rad51. We now also know that it is not involved in DNA Damage repair so we can search for other possible functions!