Jan 5, 2016

An acetoacetate-inducible bacterial sensor

BioRxiv : the Preprint Server for Biology
David T GonzalesChris P Barnes

Abstract

Whole cell biosensors have great potential to be used as diagnostic and treatment tools in biomedical applications. Bacterial biosensors that respond to specific metabolites can help analyze spatio-temporal gradients in the body or cue expression of therapeutic agents in diseased sites. In this study, we developed an acetoacetate bacterial sensor by inserting the promoter of the atoSC two-component system (TCS) into a promoterless GFP expression plasmid and transformed it into E. coli DH5α and E. coli Nissle 1917, which both contain the atoSC TCS. This bacterial biosensor has a dynamic range of 0.01-1mM of acetoacetate, which is within the range of physiological concentrations in the blood, and exhibits up to a 100-fold change of induced GFP expression as measured by relative fluorescence. Comparing combinations among the two host strains and high/low-copy plasmid variations of the biosensor, we observed the fastest and highest response to acetoacetate with the E. coli Nissle 1917 low copy plasmid biosensor. Induction experiments on the biosensor using 50mM of the short chain fatty acids (SCFA) acetate, proprionate, and butyrate showed no response, indicating its specificity between acetoacetate and SCFAs. This acetoacetate bac...Continue Reading

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Mentioned in this Paper

Study
In Vivo
Bacterial Proteins
Fatty Acids, Volatile
Body Structure
Biosensors
Promoter
Acetoacetate
Therapeutic Agent
Site

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