10 thoughts on “C65 – Nguyen

  1. You mentioned other students getting different results than you (as a cause of random or regulated DNA damage), is there a particular reason why it would increase in random but decrease in regulated?

    1. Hi Mya, thanks for your question! I actually re-did my gene expression after DNA damage experiment and got the same results as other students studying my gene, so there was decreased expression after random DNA damage occurred. Either way, different genes may be expressed in random versus regulated DNA damage because a particular gene (possibly Hyproc14) may be important during conjugation when Tetrahymena thermophila rearrange their DNA in a regulated manner. Perhaps that gene is necessary to induce double-stranded DNA breaks at a particular site. However, when it comes to random DNA damage, different genes and mechanisms may be required to repair that damage and prevent negative consequences for the organism.

  2. In the methods of how your primers for Hyproc14 were made it is mentioned that pimer3plus and BLAST are used. How do these to work together or how are they used to design the primers?

    1. Hello, we used both programs in conjunction with each other. Primer3Plus allowed us to design our primers, and it actually gave us numerous options that we could use (I recall getting 5 different options). Then we input our primers into BLAST to see if they were similar to other genes. Our goal was to get a unique set of primers that would ONLY anneal to our target gene, so we used BLAST to select primer sets that had the lowest similarity to other sequences (except the target gene) to decrease the chances of our primers annealing to and amplifying a gene that isn’t our target gene. Great question!

  3. Hi DeeDee, great presentation! Did these results you and your group got surprise you at all? Or was it clear after looking over it again? Thanks!

    1. Hi Sydney, thanks for your question! I was definitely surprised with my initial results because I expected to see at least some expression of Hyproc14 in my untreated samples, so it was strange that no band showed up. I believed that it was due to experimental error, so I actually re-did the gene expression experiment and got results that coincided with other students (decreased expression of Hyproc14 after DNA damage was induced). This didn’t match my hypothesis that Hyproc14 expression would increase, but that could just mean that this gene is important during regulated DNA damage (which occurs during conjugation) and not so important during random DNA damage.

  4. Hey great job. Do you know whether Hyproc14 is expressed more at random DNA damage or regulated DNA damage?

    1. Hi Nick, after re-doing my gene expression experiment, I ended up seeing decreased expression of Hyproc14 after random DNA damage was induced with hydroxyurea (like other students studying this gene saw). Because of that, it seems like this gene is expressed more during regulated DNA damage (conjugation). This matches with the gene expression profiles that I saw while doing background research on Hyproc14 because I saw that Hyproc14 expression peaks significantly during C14, a stage of conjugation. That’s actually how I decided to name this gene “Hyproc14.” Thanks for your question!

Leave a Reply