title: A homogeneous assay for highly sensitive detection of CaMV35S promoter in transgenic soybean by förster resonance energy transfer between nitrogen-doped graphene quantum dots and Ag nanoparticles.
abstract: In this work, a novel homogeneous assay for DNA quantitative analysis based on förster resonance energy transfer (FRET) was developed for cauliflwer mosaic virus 35s (CaMV35S) promoter of transgenic soybean detection. The homogenous FRET of fluorescence signal was fabricated by DNA hybridization with probe modified nitrogen-doped graphene quantum dots (NGQDs) and silver nanoparticles (AgNPs), which acted the donor-acceptor pairs for the first time. The highly efficient FRET and unique properties of the NGQDs made the proposed FRET system as a functionalized detection platform for labelling of DNA. Upon the recognition of specific target DNA (tDNA), the FRET between NGQDs and AgNPs was triggered to produce fluorescence quenching, which could be used for tDNA detection. The fabricated homogeneous FRET assay displayed a wide linear range of 0.1-500.0 nM and a low limit of detection 0.03 nM for the detection of CaMV35S (S/N = 3). This proposed biosensor revealed high specificity to detect tDNA, with acceptable intra-assay precision and excellent stability. This method was successfully applied to identify the real sample of 0.5% containing transgenic soybean, which achieved the most of national law regulations. This assay was further validated by polymerase chain reaction as the genetically modified organisms, suggesting that the proposed FRET system is a feasible tool for the further daily genetically modified organism detection.
authors.affiliation: School of Food and Biological Engineering, Jiangsu University, Zhenjiang, 212013, China.
authors.forename: Yaqi
authors.initials: Y
authors.lastname: Li
authors.affiliation: School of Food and Biological Engineering, Jiangsu University, Zhenjiang, 212013, China.
authors.forename: Li
authors.initials: L
authors.lastname: Sun
authors.affiliation: School of Chemistry and Chemical Engineering, Jiangsu University, Zhenjiang, 212013, China.
authors.forename: Jing
authors.initials: J
authors.lastname: Qian
authors.affiliation: School of Food and Biological Engineering, Jiangsu University, Zhenjiang, 212013, China.
authors.forename: Chengke
authors.initials: C
authors.lastname: Wang
authors.affiliation: School of Chemistry and Chemical Engineering, Jiangsu University, Zhenjiang, 212013, China.
authors.forename: Qian
authors.initials: Q
authors.lastname: Liu
authors.affiliation: School of Food and Biological Engineering, Jiangsu University, Zhenjiang, 212013, China.
authors.forename: En
authors.initials: E
authors.lastname: Han
authors.affiliation: School of Chemistry and Chemical Engineering, Jiangsu University, Zhenjiang, 212013, China.
authors.forename: Nan
authors.initials: N
authors.lastname: Hao
authors.affiliation: Sinograin Zhenjiang Grains & Oils Quality Testing Center Co., Ltd., Zhenjiang, 212013, China.
authors.forename: Liuping
authors.initials: L
authors.lastname: Zhang
authors.affiliation: School of Food and Biological Engineering, Jiangsu University, Zhenjiang, 212013, China. Electronic address: jrcai@ujs.edu.cn.
authors.forename: Jianrong
authors.initials: J
authors.lastname: Cai
authors.affiliation: School of Chemistry and Chemical Engineering, Jiangsu University, Zhenjiang, 212013, China. Electronic address: wangkun@ujs.edu.cn.
authors.forename: Kun
authors.initials: K
authors.lastname: Wang
mesh_terms: D017796:Caulimovirus / Q000235:genetics*; D031541:Fluorescence Resonance Energy Transfer; D006108:Graphite / Q000737:chemistry*; D053768:Metal Nanoparticles / Q000737:chemistry*; D046529:Microscopy, Electron, Transmission; D009584:Nitrogen / Q000737:chemistry*; D030821:Plants, Genetically Modified / Q000235:genetics / Q000821:virology*; D016133:Polymerase Chain Reaction; D011401:Promoter Regions, Genetic; D045663:Quantum Dots; D015203:Reproducibility of Results; D012834:Silver / Q000737:chemistry*; D013025:Glycine max / Q000235:genetics / Q000821:virology*; D013057:Spectrum Analysis / Q000379:methods
chemical_list: D012834:Silver; D006108:Graphite; D009584:Nitrogen
keywords: Fluorescence resonance energy transfer; Homogeneous assay; Nitrogen-doped graphene quantum dots; Silver nanoparticles; Transgenic soybean