生物荧光传感器检测环境水样中氨基甲酸酯类农药残留

A Sensitive Detection Method for Carbamate Pesticide by Fluorescence Biosensor

建立了简便、灵敏的氨基甲酸酯类农药生物荧光传感器及其检测方法。通过构建氨基甲酸酯类农药降解菌H5基因组文库，筛选出氨基甲酸酯类农药特异性响应功能基因的调控序列，将其与增强型绿色荧光蛋白基因（EGFP）连接，构建了非细胞体系生物荧光传感器H12-E，非细胞体系蛋白浓度为1．0g/L。在室温条件下，对不同浓度呋喃丹标准液进行检测，30min即可检测出龟级氨基甲酸酯类农药总量，灵敏度高于国家标准，线性范围1×10^-9～1×10^-4g/L。采集吉林省松花江流域等水样，检测氨基甲酸酯类农药残留，并向伊通河水样中添加高中低3个水平呋喃丹标准液，方法回收率95％-110％，相对标准偏差（RSD）2．4％～4．9％。本方法可望用于环境水体中氨基甲酸酯类农药残留的快速检测。

A convenient and sensitive fluorescence biosensor for carbamat pesticides was developed. By establishment of the genomic library of carhamat pesticides degrading bacteria H5, the regulatory sequence of carbamat pesticide responding gene was screened, which was then linked to gene sequence of green fluorescent protein to fabricate the cell-free fluorescence biosensor H12-E. The protein concentration of the cell-free system was 1.0 g/L. The carbofuran standards with different concentrations were detected at room temperature, and the results showed that the fg level of carbamat pesticides could be detected within 30 rain at room temperature. The sensitivity was above the national standard, and the linear range was 1 ng/L-100 μg/L. The water samples of Songhua River in Jilin province was collected and detected carbamate pesticide residues. Three levels of carbofuran were spiked into samples of Yitong River extract. The overall recoveries and the relative standard deviations （RSDs） were in the range of 95%-110% and 2.4%-4.9%. This assay proved to be a suitable tool for fast detection of carbamat pesticides residue in environmental water samples.