A paper-based, cell-free biosensor system for the detection of heavy metals and date rape drugs

Autor: Gräwe, Alexander, Dreyer, Anna, Vornholt, Tobias, Barteczko, Ursela, Buchholz, Luzia, Drews, Gila, Ho, Uyen Linh, Jackowski, Marta Eva, Kracht, Melissa, Lüders, Janina, Bleckwehl, Tore, Rositzka, Lukas, Ruwe, Matthias, Wittchen, Manuel, Lutter, Petra, Müller, Kristian, Kalinowski, Jörn
Jazyk: angličtina
Rok vydání: 2019
Předmět:
Agrobacteria
Light
Hydroxybutyrates
Gene Expression
Biosensing Techniques
Protein Synthesis
Plant Science
Biochemistry
Plant Microbiology
Physics
Electromagnetic Radiation
Chemical Synthesis
Detectors
Substance Abuse Detection
Chemistry
Artificial Light
Physical Sciences
Engineering and Technology
Medicine
Synthetic Biology
Research Article
Chemical Elements
Biosynthetic Techniques
Science
Green Fluorescent Proteins
Equipment
Research and Analysis Methods
Agrobacterium Tumefaciens
Microbiology
Metals
Heavy
DNA-binding proteins
Genetics
Humans
Gene Regulation
Communication Equipment
Cell-Free System
Bacteria
Illicit Drugs
Organisms
Biology and Life Sciences
Proteins
Regulatory Proteins
Biosensors
Rape
Metallic Mercury
Cell Phones
Water Pollutants
Chemical
Synthetic Biosensors
Transcription Factors
Zdroj: PLoS ONE, Vol 14, Iss 3, p e0210940 (2019)
PLoS ONE
ISSN: 1932-6203
Popis: Biosensors have emerged as a valuable tool with high specificity and sensitivity for fast and reliable detection of hazardous substances in drinking water. Numerous substances have been addressed using synthetic biology approaches. However, many proposed biosensors are based on living, genetically modified organisms and are therefore limited in shelf life, usability and biosafety. We addressed these issues by the construction of an extensible, cell-free biosensor. Storage is possible through freeze drying on paper. Following the addition of an aqueous sample, a highly efficient cell-free protein synthesis (CFPS) reaction is initiated. Specific allosteric transcription factors modulate the expression of 'superfolder' green fluorescent protein (sfGFP) depending on the presence of the substance of interest. The resulting fluorescence intensities are analyzed with a conventional smartphone accompanied by simple and cheap light filters. An ordinary differential equitation (ODE) model of the biosensors was developed, which enabled prediction and optimization of performance. With an optimized cell-free biosensor based on the Shigella flexneri MerR transcriptional activator, detection of 6 mug/L Hg(II) ions in water was achieved. Furthermore, a completely new biosensor for the detection of gamma-hydroxybutyrate (GHB), a substance used as date-rape drug, was established by employing the naturally occurring transcriptional repressor BlcR from Agrobacterium tumefaciens.
Databáze: OpenAIRE
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