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10 thoughts on “C49 – Ogden”
What else can explain the upregulation of Nuftr1 during conjugation?
Good question! Nuftr1 could also be involved in conjugation in other capacities, such as the process of damaging DNA or binding to DNA simply to replicate it. Nuftr1 is hypothesized to bind to DNA, and if that’s so it could have a lot of roles in regulating DNA in other ways than just repairing it!
How did you “expect” certain bands
Awesome question! We expect certain band lengths just based on the length of the DNA that will be amplified with the specific primers, for example my gene’s primers would amplify a sequence of DNA that is 713 bp long. We except band’s appearance based on what type of control they are, so negative controls are expected to have no bands and if they do have bands, that means there is some contamination etc. Rad51 was expected to have an up regulation after damage because it is known to help repair DNA during conjugation, while Ftt18 was a loading control and expected to have bands of the same brightness because it is equally expressed with or without damage.
Good job on your poster. What other cellular functions does Nufter 1 have?
Great question! Almost none of these T. thermophila genes have been studied really at all, so their functions are largely unknown. We only know Nuftr1’s gene expression profile from experiments analyzing expression of all of the Tetrahymena genes. However, we do know that Nuftr1 has a true homolog in Tetrahymena borealis that is a High Mobility Group (HMG)-box protein that has a role in binding to DNA. Therefore, we suspect that it Nuftr1 could bind to DNA, we just don’t know why or how.
Do you think you would have similar or different results if you repeated the trials using a Northern Blot instead?
Good question! Because with RT-PCR we had to add an extra step of translating the mRNA to cDNA, some errors or differing effects could have occurred during that time. Northern blotting gets rid of that extra step because it analyzes mRNA directly, therefore possibly reducing any errors that could have arisen converting mRNA to cDNA.
Could Nuftr1 be molecularly modified and experimentally tested to try and find a specific structure that would show more bands on the gel?
How can using quantitative PCR affect the results of Nufter1 gene expression?