Protein Immobilization Research Weeks 1-2

Within the first two weeks of working in the research lab, I think I’ve finally begun to understand the meaning of the phrase “learning curve.” Even though I had been coming in a few hours each week the past two semesters, being in the lab from 9-5 working on your own project is a much different experience. It involves more independence, self-reliance, and willingness to troubleshoot. Vastly different from the teaching labs, the experiments I’ve undertaken rely much more on my own competence, adding both pressure and excitement to the research experience. I am incredibly appreciative of both Dr. Young and the senior members in the lab for their patience in answering all my questions. Ironically, the farther along I get in my research, the more I realize there is to discover. Everyday I learn something new, and come home exhausted from throwing my entire mental energy into the day.

In my research, I’ve been working on trying to express the mutants of the bacteria SSO. Building on the work of previous lab members, I am trying to maximize expression of both the wild type SSO, encoded into a new pET plasmid, as well as mutate and express SSO proteins with an unnatural amino acid (UAA). Young’s lab has previously encoded an unnatural amino acid into a marker protein by replacing a TAG stop codon with their UAA. My aim will be to accomplish this with the much more functional SSO protein, for hopeful use in biosensors and biochips. The next steps in my project are to express our current mutants, and run an assay to prove functionality. I will also examine the protein DNA sequence to determine if there are other viable mutant possibilities.While I have run into multiple roadblocks, I’m hopeful with time and hard work I will be able to express this protein and fulfill my goal for the summer.

I’m also working on further optimizing the Glaser-Hay reaction our lab has previously published work on, as well as other multi-molecule coupling reactions. An idea I am especially excited about is the incorporation of a thiol into an alkyne created by a Glaser-Hay reaction. This reaction, relatively easy to undertake, has been proven successful with other proteins treated with irradiation and a photoactivator molecule. A successful product from this reaction would have huge implications for the medical community, as it could  revolutionize drug delivery molecules, such as coupling an antibiotic to a drug seeking protein.

In conclusion, I have lots to work on for the remainder of the summer, and am eager to see what unfolds. I have already learned so much in such a short period of time, making the next 8 weeks even more exciting. With hard work, collaboration with my peers, and Dr. Young’s guidance, I’m hopeful my research will lead to advances in our lab, and our work with unnatural amino acids.

 

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