Coupling High-Performance Thin-Layer Chromatography with Bacterial Genotoxicity Bioreporters.
Dror ShakibaiCarolin RiegrafLiat MoscoviciGeorg ReifferscheidSebastian BuchingerShimshon BelkinPublished in: Environmental science & technology (2019)
We present an innovative technological platform for monitoring the direct genotoxicity of individual components in complex environmental samples, based on bioluminescent Escherichia coli genotoxicity bioreporters, sprayed onto the surface of a high-performance thin-layer chromatography (HPTLC) plate. These sensor strains harbor plasmid-borne fusions of selected gene promoters of the E. coli SOS DNA repair system to the Photorhabdus luminescens luxABCDE gene cassette, and mark by increased luminescence the presence of potentially DNA-damaging sample components separated on the plate. We demonstrate an "on plate" quantifiable dose-dependent response to several model genotoxicants (without metabolic activation). We further demonstrate the applicability of the system by identifying as genotoxic specific components of HPTLC-separated influent and effluent samples of wastewater treatment plants, thereby alleviating the need for a comprehensive chemical analysis of the sample.
Keyphrases
- wastewater treatment
- escherichia coli
- dna repair
- mass spectrometry
- antibiotic resistance genes
- high speed
- dna damage
- copy number
- tandem mass spectrometry
- genome wide
- liquid chromatography
- oxide nanoparticles
- biofilm formation
- genome wide identification
- klebsiella pneumoniae
- high performance liquid chromatography
- dna damage response
- high throughput
- oxidative stress
- circulating tumor
- gene expression
- single molecule
- quantum dots
- transcription factor
- microbial community
- high resolution
- single cell
- crispr cas
- risk assessment
- climate change
- human health
- energy transfer
- ms ms
- circulating tumor cells
- solid phase extraction