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A next-generation bacterial biosenso...
~
Martineau, Rhett Landen.
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A next-generation bacterial biosensor for remote or automated applications.
Record Type:
Language materials, printed : Monograph/item
Title/Author:
A next-generation bacterial biosensor for remote or automated applications./
Author:
Martineau, Rhett Landen.
Description:
244 p.
Notes:
Source: Dissertation Abstracts International, Volume: 69-03, Section: B, page: 1766.
Contained By:
Dissertation Abstracts International69-03B.
Subject:
Biology, Microbiology. -
Online resource:
http://pqdd.sinica.edu.tw/twdaoapp/servlet/advanced?query=3304861
ISBN:
9780549513056
A next-generation bacterial biosensor for remote or automated applications.
Martineau, Rhett Landen.
A next-generation bacterial biosensor for remote or automated applications.
- 244 p.
Source: Dissertation Abstracts International, Volume: 69-03, Section: B, page: 1766.
Thesis (Ph.D.)--Arizona State University, 2008.
The performance of the dual-reporter construct is compared with conventional recA::gfp and the newer recA:: mCherry. Mitomycin C (MMC) is used as a model inducer of DNA damage to modulate recA activity in Escherichia coli hosts.
ISBN: 9780549513056Subjects--Topical Terms:
1017734
Biology, Microbiology.
A next-generation bacterial biosensor for remote or automated applications.
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244 p.
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Source: Dissertation Abstracts International, Volume: 69-03, Section: B, page: 1766.
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Thesis (Ph.D.)--Arizona State University, 2008.
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The performance of the dual-reporter construct is compared with conventional recA::gfp and the newer recA:: mCherry. Mitomycin C (MMC) is used as a model inducer of DNA damage to modulate recA activity in Escherichia coli hosts.
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Many current methods of bacterial biosensing are based on optical techniques using high-performance benchtop instrumentation. For remote, distributed applications including environmental hazard assessment and space biology, however, these systems are not ideal due to high power consumption, bulkiness, and complexity.
520
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This work demonstrates that sophisticated biological assays can be conducted using simpler optical systems more suitable for remote applications. A compact detector comprised of close-coupled light-emitting diodes (LEDs) and simple photodiodes (PDs) is used to quantify two-color fluorescence and cell biomass in complex mixtures. The performance of this system supports new paradigms for remote biosensing.
520
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A reporting construct is presented which harnesses biological mechanisms for signal amplification and takes advantage of multi-spectral detection capabilities. A simple regulatory architecture is presented whereby induction of the Recombinase A (recA) promoter in response to deoxyribonucleic acid (DNA) damage increases both the intracellular content of green fluorescent protein (GFP) and the lactose metabolism repressor (LacI). An amplification stage is comprised of a hybrid tryptophan-lactose (tac) promoter, repressed by LacI but otherwise expressive, with transcriptional control over a red fluorescent protein (RFP), mCherry. Thus, DNA damage results in an increase of GFP and an amplified decrease in RFP.
520
$a
Single-reporter RFP constructs outperformed GFP constructs in the miniature detector, achieving a limit of detection better than 1 nM MMC. In 2 of 3 triplicate experiments using the dual reporter construct, 0.1 nM MMC concentrations are discernible from no-treatment samples in the compact optical system. Thus, the lowest concentrations of MMC linked to DNA damage by the classic Ames' test can be sensed using compact and inexpensive optics. Given that recA responds to numerous genotoxins including radiation sources, sensitivity at this level suggests the potential for remote monitoring applications. These results demonstrate that increasing the sophistication of biological assays can improve overall system gain and signal to noise ratio such that simpler optical systems can be used for monitoring in remote applications.
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School code: 0010.
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Arizona State University.
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http://pqdd.sinica.edu.tw/twdaoapp/servlet/advanced?query=3304861
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