William Wittwer and Dr. Laura Bridgewater, Microbiology
One of the main things that I have learned over the course of this project is that things work better in theory than in practice. I spent two semesters attempting to builda plasmid containing the the p17 cDNA sequence in order to test the ability of p17 as an activator of the Coll11a2 gene. Unfortunately, I was not able to produce the plasmid given to me.
We received the original p17 cDNA in a plasmid from another university. I could not use the protein as it was sent to us however because of the fact that I needed a plasmid which could be expressed in a eukaryotic cell, whereas the plasmid the cDNA was sent in was a prokaryotic expression plasmid. What should have been a simple excision and ligation into the correct plasmid, never wanted to function. I was able to successfully remove the desired DNA from the plasmid that was sent to us, but whenever I attempted to ligate the plasmid into a eukaryotic expression plasmid I met with failure.
Throught the course of experiment after experiment I determined that the problem was not with our restriction enzymes used to cut the plasmids but with our expression plasmid. For some reason the plasmid that we purchased was not as advertised. It contained more that one EcoRI restriction sites where it had been advertised as containing only one. Unfortunately by the time I was able to figure this out, time had run out on me and both semesters had passed.
Because of this inability of mine to create the plasmid that was necessary to perform my project I was not able to present my research because I was not able to carry it out. I do not feel that this was wasted time however. I was able to learn the effort that goes behind each part of the research process. I was able to see how an excellent professor interacts with students attempting to perform in a laboratory setting. I was able to obtain hands on experience in a lab which I hope will lead to more research in the future.
I am very grateful for ORCA and BYU in helping me to grow and to learn.