Rachel Siebenmorgen with her gel electrophoresis product.
Author: Rachel Siebenmorgen | Major: Biology
During the Summer 2021 term, I started my research with Dr. Mack Ivey of Biological Sciences in his research lab. As a student majoring in Biology and minoring in Medical Humanities and Spanish, I have found this to be a very rewarding and interesting experience and I am excited to continue my research into the Fall 2021 term. My thesis topic is focused on the opportunistic pathogen well known in the medical field called Clostridium difficile. Within the healthcare setting, specifically long-term care, C. difficile has shown to be the primary cause of infectious diarrhea and colitis, therefore, it is an important organism to study. Coming into the Summer 2021 term, I had produced three-dimensional protein structures using ab initio modeling programs. One of the goals of my research is to test the accuracy of my models using various experimental techniques in the lab. To contribute to this goal, I was able to get comfortable in the lab by doing multiple sessions of PCR (polymerase chain reaction) tests and gel electrophoresis tests to confirm the stability of various DNA samples and PCR master mixes. The future plan of our research is to further explore the linker region of the SpoIIE protein within C. difficile using the three-dimensional protein structures previously mentioned and other lab processes such as cloning, fluorescence assay, and immunoprecipitation (IP) processes.
In the healthcare field, the well-being of patients takes top priority, however, problems arise when pathogens such as C. difficile are presented an opportunity to attack susceptible hosts. The normal human body has intestines flourishing with normal flora that help keep us healthy, however, a problem occurs when these bacteria are decreased or eliminated completely due to antibiotic therapy. The absence of flora in the gut creates an unintentional optimal growing environment for this gram positive, endospore-forming, obligate anaerobe. C. difficile attacks the intestinal tract by releasing two toxins, enterotoxin A and cytotoxin B, or also known as TcdA and TcdB, which are responsible for the sporulation and infection process causing chronic diarrhea, pseudomembranous colitis, and several other clinical features. Due to this, the research being conducted in the Ivey lab is especially pertinent to all patients and healthcare workers.
This past summer of research was rewarding particularly through experience as I am now more comfortable in the lab and know how to run PCR and gel electrophoresis very well. Upon doing my first round of PCR and gel electrophoresis, it was discovered that either the master mix or DNA sample had gone bad. To test this, another round of PCR and gel was completed using a supplement added to the mixture and thus it was determined the master mix was the culprit. Finally, another round of PCR and gel was completed with new primers which would ensure optimal DNA replication. It was necessary to complete these processes and others not listed to ensure everything was working as it should when using for the more advanced tests. While it was planned to get further into the research project this summer, things did not work out as planned. This helped me to learn that sometimes unexpected obstacles will appear that may stunt my progress, but so long as I continue, I will be happy with what I accomplished. In addition, I am so much more eager to get back into the lab and learn new lab processes and complete the more advanced tests needed. I also was able to order a white lab coat so that is especially exciting.
The process of finding my way into Dr. Ivey’s lab was not as unpersonal as looking through the biology lab directory and choosing one based on a blurb. I was drawn to Dr. Ivey’s lab through a graduate student who was a TA for my honors Microbiology lab who recommended I send him an email to see if there were any open positions and lucky for me there was. I chose to work on the C. difficile research project as I aspire to a pharmacist in the near future, and I know learning about what could affect my future patients would benefit not only them, but me as well. Throughout my time working with Dr. Ivey, he has played a vital role as my mentor by providing ample support, guidance, and knowledge. I am excited to pursue further research within the Ivey lab and I am grateful to the Honors College Research Grant for helping to fund it.