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Bacterial sensors synthetic design and application principles
Author
Title
Bacterial sensors [electronic resource] : synthetic design and application principles / Jan Roelof van der Meer.
ISBN
9781598299120 (electronic bk.)
9781598299113 (pbk.)
Published
San Rafael, Calif. (1537 Fourth Street, San Rafael, CA 94901 USA) : Morgan & Claypool, c2011.
Physical Description
1 online resource (ix, 153 p.) : ill., digital file.
Local Notes
Access is available to the Yale community.
Notes
Part of: Synthesis digital library of engineering and computer science.
Series from website.
Access and use
Access restricted by licensing agreement.
Summary
Bacterial reporters are live, genetically engineered cells with promising application in bioanalytics. They contain genetic circuitry to produce a cellular sensing element, which detects the target compound and relays the detection to specific synthesis of so-called reporter proteins (the presence or activity of which is easy to quantify). Bioassays with bacterial reporters are a useful complement to chemical analytics because they measure biological responses rather than total chemical concentrations. Simple bacterial reporter assays may also replace more costly chemical methods as a first line sample analysis technique. Recent promising developments integrate bacterial reporter cells with microsystems to produce bacterial biosensors. This lecture presents an in-depth treatment of the synthetic biological design principles of bacterial reporters, the engineering of which started as simple recombinant DNA puzzles, but has now become a more rational approach of choosing and combining sensing, controlling and reporting DNA 'parts'. Several examples of existing bacterial reporter designs and their genetic circuitry will be illustrated. Besides the design principles, the lecture also focuses on the application principles of bacterial reporter assays.A variety of assay formats will be illustrated, and principles of quantification will be dealt with. In addition to this discussion, substantial reference material is supplied in various Annexes.
Other formats
Also available in print.
Format
Books / Online
Language
English
Added to Catalog
April 25, 2013
Series
Synthesis lectures on synthetic biology, # 2
System details note
Mode of access: World Wide Web.
System requirements: Adobe Acrobat Reader.
Bibliography
Includes bibliographical references.
Contents
1. Short history of the use of bacteria for biosensing and bioreporting
Early warning systems
Early use of bacterial 'bioreporters'
References
2. Genetic engineering concepts
Introduction to genetic sensing/-reporting circuits
Central idea
Intercept design
Orthogonal design
Design parts
Use of transcriptional activators
Choice of regulatory proteins
The XylR/DmpR family of transcription activators
The HbpR system
Use of transcriptional repressors
Regulators from heavy metal resistance
The MerR system
ArsR-based designs for arsenic detection
Network interception designs
General motivation
SOS response network intercept design
Promoter engineering
General notions
Promoter engineering in the Ars system
Physiological control of the XylR-regulated Pu promoter
Dual responsive control switches
Directed evolution of promoters
Response heterogeneity in populations
Engineering new effector specificities
General concept
Effector domain mutagenesis in XylR/DmpR-type proteins
Effector binding pocket modeling
Aptamers
Complex signal-transduction chains
General concept
Periplasmic binding proteins and phosphotransfer relay
Multinode networks
Logic gates, transcriptional noise, amplification
Reporter proteins
Choice and specificities of common reporter proteins
Reporter gene vectors
Embedding of sensing/-reporting circuits in a cellular chassis
References
3. Measuring with bioreporters
Assay principles
Relative and absolute measurements
End-point and kinetic measurements, measurement transformations
Population measurements
Spiking
Theory of analyte provision and transport
Calculation of compound concentrations in reporter cells
Non-diffusive and non-conservative bioreporters
Concept of bioavailability and bioaccessibility
Bioavailable fractions
Bioavailability and bioaccessibility reporter measurements
Bioreporter assay types
Aqueous assays
Gas phase measurements
Solid phase assays
Method detection limits, accuracy and precision
References
4. Epilogue
Summary
Future directions
References
A. Bacterial bioreporter designs targeting organic compounds
References
B. Bacterial bioreporter designs targeting (heavy) metals and metalloids
References
C. Bacterial bioreporter designs responsive to toxicity or stress conditions
References
D. Example bioreporter protocols
Quantitative arsenite measurements with an E. coli LuxAB Luciferase bioreporter
References
Arsenic measurement using an E. coli GFP biosensor by epifluorescence microscopy
References
Arsenic measurements with an E. coli beta-galactoside bioreporter
References
Sample pretreatment
Water
Rice
References
Author's biography.
Subjects (Medical)
Biosensing Techniques.
Genes, Reporter.
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