In this project, we engineered two Lactic Acid Bacteria species, Lactococcus lactis and Lactobacillus johnsonii, for maintaining healthy homeostasis of the vaginal flora and achieving effective prevention of menstruation-related Toxic Shock Syndrome. In this process, we designed and verified the activity of multiple biobricks parts, created a new prokaryotic promoter strength analysis program, and executed 6 engineering cycles.
We created various new genetic parts, specially designed for functioning in Lactic Acid Bacteria and compatible with Biobricks Assembly. We identified strong Lactococcus promoters such as nisA promoter and PTSIIC-celA promoter, and verified their function with experiments. We also designed a CRISPR/Cas9 plasmid specifically for Lactococcus. For detailed information, please see our Parts Registry.
We created a Prokaryotic (Motif-Based) Promoter (Strength Comparison) Analysis Program (PPAP). It was developed specifically for Lactococcus lactis subsp. lactis IL1403 using regulatory sites data obtained from the RegPrecise database: https://regprecise.lbl.gov/, and can be optimised for any species whose data is available on the same site. For detailed information, please see PPAP Design.
We completed 6 DBTL Cycles, where we successfully predicted and tested the activities of 14 Lactococcus lactis promoters, verified construction of the nisRK enhancer system for the Lactic Acid Bacteria nisA promoter, and tested an H2O2 overexpression system in Lactobacillus johnsonii. For detailed information, please see our Engineering Page.
Our lab notebook documented all our experimental protocols and preliminary data, which evidences our contributions and could potentially provide insight for future research. Please see Lab Notebook for details.
