Antibiotic resistance is a serious, and rapidly growing global threat. Here, we will design a sensitive screening method to detect antibiotic resistance genes (ARGs) in bacteria using a graphene oxide-based biosensor utilizing engineered zinc finger proteins (ZFPs). Our approach relies on the on and off effect of fluorescence signal in the absence and presence of target ARGs, respectively. Two-dimensional graphene oxide (GO) sheet possesses unique electronic, thermal, and mechanical properties. The quenching ability of GO creates novel methods for detection of biomolecules. Quantum dot-labeled DNA-binding zinc finger proteins (ZFP) can bind to GO via stacking interactions of aromatic and hydrophobic residues in conjunction with hydrogen bonding interaction between hydroxyl or carboxyl groups of GO and hydroxyl or amine groups of the protein. Due to fluorescence resonance energy transfer (FRET) between quantum dot (QD) and GO when they are in close proximity, fluorescence signal of QD-labeled ZFP will be quenched. In the presence of target DNA, the bound DNA-protein complex is released from GO, restoring the fluorescence signal. Here, we optimized GO concentrations used in the assay and QD-labeling on ZFPs and determined the sensitivity of our system.

Developing a new sensing technology for detecting antibiotic resistance genes utilizing engineered zinc finger proteins and graphene oxide

2019 Spring National Meeting

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