title: A novel recombinant cell fluorescence biosensor based on toxicity of pathway for rapid and simple evaluation of DON and ZEN.
abstract: During an exposure, humans and animals are most often exposed to a mixture rather than individual mycotoxins. In this study, a Human Embryonic Kidney 293 cell (HEK-293) fluorescence sensor was developed to detect and evaluate mycotoxins, deoxynivalenol (DON) and zearalenone (ZEN) compounds, produced by Fusarium culmorum that are common food contaminants. TRE-copGFP (green fluorescent protein) and ERE-TagRFP (red fluorescent protein) plasmids were constructed and cotransfected into HEK-293 cells through a highly efficient, lipid-mediated, DNA-transfection procedure. Results show that fluorescence intensity was proportional to DON and ZEN concentrations, ranging from 2 to 40 ng/mL and 10 to 100 ng/mL respectively, with a detection limit of 0.75 ng/mL and 3.2 ng/mL respectively. The EC50 of DON and ZEN are 30.13 ng/mL and 76.63 ng/mL respectively. Additionally, ZEN may have a synergistic effect on enhancing AP-1 activity of the toxicity pathway of DON. These data indicate the high sensitivity and effectiveness of our biosensor system in the evaluation of the combined toxicity of ZEN, DON and their derivatives. In addition, this approach is suitable for an early warning method for the detection of ZEN and DON family mycotoxins contamination without higher-priced, conventional analytical chemistry methods.
authors: State Key Laboratory of Food Science and Technology, School of Food Science of Jiangnan University, School of Food Science Synergetic Innovation Center of Food Safety and Nutrition, Wuxi, Jiangsu, 214122, China.
authors: Jian
authors: J
authors: Ji
authors: State Key Laboratory of Food Science and Technology, School of Food Science of Jiangnan University, School of Food Science Synergetic Innovation Center of Food Safety and Nutrition, Wuxi, Jiangsu, 214122, China.
authors: Wenshu
authors: W
authors: Gu
authors: State Key Laboratory of Food Science and Technology, School of Food Science of Jiangnan University, School of Food Science Synergetic Innovation Center of Food Safety and Nutrition, Wuxi, Jiangsu, 214122, China.
authors: Chao
authors: C
authors: Sun
authors: State Key Laboratory of Food Science and Technology, School of Food Science of Jiangnan University, School of Food Science Synergetic Innovation Center of Food Safety and Nutrition, Wuxi, Jiangsu, 214122, China.
authors: Jiadi
authors: J
authors: Sun
authors: State Key Laboratory of Food Science and Technology, School of Food Science of Jiangnan University, School of Food Science Synergetic Innovation Center of Food Safety and Nutrition, Wuxi, Jiangsu, 214122, China.
authors: Hui
authors: H
authors: Jiang
authors: State Key Laboratory of Food Science and Technology, School of Food Science of Jiangnan University, School of Food Science Synergetic Innovation Center of Food Safety and Nutrition, Wuxi, Jiangsu, 214122, China.
authors: Yinzhi
authors: Y
authors: Zhang
authors: State Key Laboratory of Food Science and Technology, School of Food Science of Jiangnan University, School of Food Science Synergetic Innovation Center of Food Safety and Nutrition, Wuxi, Jiangsu, 214122, China.
authors: Xiulan
authors: X
authors: Sun
mesh_terms: D015374:Biosensing Techniques; D002469:Cell Separation; D004247:DNA / Q000737:chemistry; D005434:Flow Cytometry; D005506:Food Contamination / Q000032:analysis*; D005670:Fusarium / Q000737:chemistry; D049452:Green Fluorescent Proteins / Q000737:chemistry; D057809:HEK293 Cells; D006801:Humans; D008055:Lipids / Q000737:chemistry; D008164:Luminescent Proteins / Q000737:chemistry; D009183:Mycotoxins / Q000032:analysis*; D017382:Reactive Oxygen Species / Q000737:chemistry; D011994:Recombinant Proteins / Q000737:chemistry*; D014162:Transfection; D014255:Trichothecenes / Q000032:analysis*; D015025:Zearalenone / Q000032:analysis*; D000097573:Red Fluorescent Protein
chemical_list: D004247:DNA; D049452:Green Fluorescent Proteins; D008055:Lipids; D008164:Luminescent Proteins; D009183:Mycotoxins; D017382:Reactive Oxygen Species; D011994:Recombinant Proteins; D014255:Trichothecenes; D015025:Zearalenone; D000097573:Red Fluorescent Protein; C007262:deoxynivalenol