Project Description

Heterogeneity of cell populations caused by quorum sensing leads to variability in gene expression that is hard to predict. During biomanufacturing, elevating protein expression will lead to gain of profit. Decreasing protein expression can also be beneficial in situations where undesirable biofilms may form on medical equipment or controlling virulence in bacteria. We will modify the existing bacteria Autoinducer-2 (AI-2) quorum sensing system by upregulating AI-2 synthesis and excretion coupled with AI-2 uptake and phosphorylation after the initial AI-2 threshold concentrations have been reached to reduce variability in induced gene expression. A model is used to predict the impacts of manipulating the expression of quorum sensing genes and guide design and experimentation. DNA assembly of a Biobrick containing pLsr, T7RPol, LsrK, LsrACDB, LuxS, YdgG, and superfolding Green Fluorescent Protein (sfGFP) enables the quantification of what percent of cells are expressing the synthetic plasmid and their levels of protein production. This device will improve the viability of autoinduction as an induction method in industries such as biomanufacturing by decreasing the variability of a cell phenotypes within a single culture to reduce costs leading to an increase in profits.