Presenter: Julissa van Renselaar, College of Engineering, Biological Engineering
Authors: Julissa van Renselaar
Faculty Advisor: Ryan Jackson, College of Science, Chemistry and Biochemistry
Institution: Utah State University
The GEAR UP lab at Utah State University (USU) was tasked with making a Bacillus subtilis classroom kit that would be safe and contained for student access. Bacillus subtilis is a sporulating bacteria that allow it to survive unfavorable conditions. A common method for disposal of endospores is autoclaving with conditions set at 121° C and 15 psi. These conditions are not favorable or accessible in a classroom setting. Proactive measures were taken by creating another condition of growth for Bacillus subtilis 168. In order to create this additional condition, a double knockout of the thymine coding genes was performed modeled after Hosseini et. al thymineless death paper. With the adjustments of protocols for cell competence, transformation, and minimal media BKE 17680 ΔthyA:: erm ΔthyB:: kan was successfully created. This double knockout strain can now be used safely in a classroom setting.Further research utilizing the Cre plasmid, pDRR244 needs to be done to loop out the antibiotic cassettes as it will affect downstream processing.