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Project Description

Heterogeneity of cell populations caused by quorum sensingThe ability to detect and to respond to cell population density by gene regulation.undefined leads to variability in gene expression that is hard to predict. During biomanufacturing, elevating quorum-induced protein expression will lead to gain of profit. Decreasing this expression can also be beneficial in situations where undesirable biofilmsAggregate of microorganisms in which cells that are frequently embedded within a self-produced matrix of extracellular polymeric substances (EPSs) adhere to each other and/or to a surface.undefined may form on medical equipment or controlling virulenceProvides a quantitative measure of the pathogenicity or the likelihood of a microbe to cause disease.undefined in bacteria. We will modify the existing bacterial quorum sensing system controlled by the Lsr operonGenes that regulate other genes responsible for protein synthesis. undefined by upregulating the synthesis and excretion of Autoinducer-2A member of a family of signaling molecules used in quorum sensing.undefined , a universal quorum molecule. This will increase population-scale AI-2 intake and phosphorylationA biochemical process that involves the addition of phosphate to an organic compound. undefined 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 Biobricks containing pLsrA promoter of the Lsr Operon.undefined , T7 RNA PolymeraseA RNA polymerase from the T7 bacteriophage that catalyzes the formation of RNA from DNA in the 5'→ 3' direction.undefined , LsrKA1-2 kinase, which catalyzes the phosphorylation of A1-2 to phospho-AI-2.undefined , LsrACDBImport protein for A1-2.undefined , LuxSAn enzyme closely linked to AI-2 production.undefined , YdgGA protein that enhances transport of AI-2.undefined , and superfolding Green Fluorescent Protein (sfGFPSuper folding green flourescent protein that is commonly used for protein expression levels.undefined ) enables both the enhancement of natural quorum sensing and the quantification of protein activation among the bacteria in a colony. This device will improve the viability of autoinductionThe induction of something by itself or without external stimuli.undefined as an induction method in industries such as biomanufacturing by decreasing the variability of a cell phenotypes and increasing expression within a single culture to reduce costs leading to an increase in profits. In addition, by insertion of T7 polymerase into a quorum-sensitive region of the genome of E. coli, we will create a chassis with specific, customizable quorum response for engineering and scientific applications.