Revision as of 19:25, 20 August 2018

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 biofilmsA protective, adhesive matrix of polymers typically produced after quorum activation.undefined may form on medical equipment or controlling virulenceThe likelihood of a microbe to cause disease based on its phenotypic state.undefined in bacteria. We will modify the existing bacterial quorum sensing system controlled by the Lsr operonLuxS Regulated (Lsr) operon responsive to AI-2.undefined by upregulating the synthesis and excretion of Autoinducer-2A universal signaling molecule used by microorganisms to coordinate group behavior through quorum sensing.undefined , a universal quorum molecule. This will increase population-scale AI-2 intake and phosphorylation The addition of a phosphate group 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 pLsrThe bidirectional promoter of the Lsr Operon.undefined , T7 RNA PolymeraseAn RNA polymerase from the T7 bacteriophage is highly selective for the pT7 promoter.undefined , LsrKAI-2 kinase, which catalyzes the phosphorylation of A1-2 to phospho-AI-2.undefined , LsrACDBActive import protein for AI-2.undefined , LuxSAn enzyme closely linked to the production of AI-2.undefined , YdgGActive export protein for AI-2.undefined , and sfGFPsuperfolding Green Fluorescent Proteinundefined 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 activation of a phenotype 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.