This study focuses on engineering whole cell-based biosensors for heavy metal detection. Through the exploitation of metalloregulatory proteins, fabrication of metal ion-responsive biosensors is achieved. Metalloregulatory proteins of the SmtB/ArsR family including arsenite-responsive ArsR, cadmium-responsive CadC, zinc-responsive CzrA, and nickel-responsive NmtR were evaluated as biosensor sensing modules. Characterization of these four metal sensing modules was accomplished through quantification of a reporter green fluorescence protein (gfp) gene. As such, biosensors pCTYC-r34ArsR-pL(ArsOvN)GFP and pCTYC-r34CadC-pL(CadOv1)GFP displayed excellent gfp expression and sensitivity to As(III) and Cd (II), respectively. These two biosensors were consequently selected and successfully implemented in soil bacterium Pseudomonas putida. Lastly, a proof of concept arsenite-responsive genetic toggle switch is proposed utilizing PurRcelR467 (PC47), a cellobiose-responsive gene, and an LAA degradation tag. Overall, this study expands the bank of metalloregulatory bioparts for heavy metal sensing in the aim of constructing an optimized water monitoring system.
Identifer | oai:union.ndltd.org:unt.edu/info:ark/67531/metadc1944355 |
Date | 05 1900 |
Creators | Draeger, Alison |
Contributors | Chan, Clement, Lee, Moo-Yeal, Yang, Yong |
Publisher | University of North Texas |
Source Sets | University of North Texas |
Language | English |
Detected Language | English |
Type | Thesis or Dissertation |
Format | Text |
Rights | Public, Draeger, Alison, Copyright, Copyright is held by the author, unless otherwise noted. All rights Reserved. |
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