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[一种用于检测农田土壤中有机磷农药甲基对硫磷的全细胞生物传感器的研制]

[Development of a whole-cell biosensor for detecting organophosphorus pesticide methyl parathion in the farmland soil].

作者信息

Ma Zhao, Li Meng

机构信息

Archaeal Biology Center, Institute for Advanced Study, Shenzhen University, Shenzhen 518061, Guangdong, China.

Shenzhen Key Laboratory of Marine Microbiome Engineering, Institute for Advanced Study, Shenzhen University, Shenzhen 518061, Guangdong, China.

出版信息

Sheng Wu Gong Cheng Xue Bao. 2023 Jul 25;39(7):2706-2718. doi: 10.13345/j.cjb.230129.

Abstract

The evaluation of the bioavailability of pollutants in soil is crucial to accurately assess the pollution risk, and whole-cell biosensor is one of the important tools for such evaluation. This study aimed to develop a novel whole-cell biosensor for the detection of methyl parathion in soil using. First, a whole-cell biosensor was constructed by the screened methyl parathion hydrolase gene, the existing specific induction element R, and the pUC19 plasmid skeleton. Then, the detection method of methyl parathion in soil extracts was established using 96-well microtiter plate as carrier and five whole-cell biosensors as indicator. The method was applied in the detection of methyl parathion in tested and field soil extracts. Taking . DH5α/pMP-AmilCP with the best detection performance as an example, this biosensor had a detection limit of 6.21-6.66 µg/L and a linear range of 10-10 000 µg/L for methyl parathion in four soil extracts. . DH5α/pMP-RFP and . DH5α/pMP-AmilCP methods have good detection performance for the analysis of methyl parathion in soil extract samples. This biosensor method can help to quickly assess the bioavailability of methyl parathion in soil, and thus help to understand the risk of soil pollution caused by organophosphorus pesticide methyl parathion.

摘要

评估土壤中污染物的生物有效性对于准确评估污染风险至关重要,而全细胞生物传感器是进行此类评估的重要工具之一。本研究旨在开发一种用于检测土壤中甲基对硫磷的新型全细胞生物传感器。首先,利用筛选出的甲基对硫磷水解酶基因、现有的特异性诱导元件R和pUC19质粒骨架构建全细胞生物传感器。然后,以96孔微量滴定板为载体、五个全细胞生物传感器为指示剂,建立土壤提取物中甲基对硫磷的检测方法。该方法应用于测试土壤提取物和田间土壤提取物中甲基对硫磷的检测。以检测性能最佳的DH5α/pMP-AmilCP为例,该生物传感器对四种土壤提取物中甲基对硫磷的检测限为6.21 - 6.66 µg/L,线性范围为10 - 10 000 µg/L。DH5α/pMP-RFP和DH5α/pMP-AmilCP方法对土壤提取物样品中甲基对硫磷的分析具有良好的检测性能。这种生物传感器方法有助于快速评估土壤中甲基对硫磷的生物有效性,从而有助于了解有机磷农药甲基对硫磷造成的土壤污染风险。

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