Master of Science (MS)
Semester of Degree Completion
Steven L. Daniel
Oxalobacter formigenes is an anaerobic, oxalate degrading organism that colonizes in the mammalian intestinal tract. Currently, O. formigenes is divided into two groups via PCR of the oxc gene. Pulsed field gel electrophoresis (PFGE) is a technique used to resolve large pieces of DNA. Typically, PFGE is used to identify differences between strains for pathogenic bacteria, but this study sought to use it to further understand the diversity of a probiotic organism. This study aimed to develop a methodology for utilizing PFGE to examine O. formigenes. The methodology involved lysing O. formigenes cells in plugs, first in proteinase K, followed by lysozyme. The cells were digested using XbaI and SacI, and run on a BioRad CHEF Mapper®. While many of the methods tested failed to produce results, attempting the two step method with proteinase K and lysozyme, resulted in bands that became clearer and were able to be analyzed. Differences were observed between strains of O. formigenes. Even strains from the more conserved group I showed variation in their DNA fingerprint using this method.
Shaw, Karen, "Diversity of the Beneficial Bacterium Oxalobacter formigenes Isolated from the Human Gut" (2016). Masters Theses. 2477.