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Application of DNA Hybridization Biosensor as a Screening Method for the Detection of Genetically Modified Food Components

机译:DNA杂交生物传感器作为转基因食品成分检测的筛选方法

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An electrochemical biosensor for the detection of genetically modified food components is presented. The biosensor was based on 21-mer single-stranded oligonucleotide (ssDNA probe) specific to either 35S promoter or nos terminator, which are frequently present in transgenic DNA cassettes. ssDNA probe was covalently attached by 5′-phosphate end to amino group of cysteamine self-assembled monolayer (SAM) on gold electrode surface with the use of activating reagents – water soluble 1-ethyl-3(3′-dimethylaminopropyl)-carbodiimide (EDC) and N-hydroxy-sulfosuccinimide (NHS). The hybridization reaction on the electrode surface was detected via methylene blue (MB) presenting higher affinity to ssDNA probe than to DNA duplex. The electrode modification procedure was optimized using 19-mer oligoG and oligoC nucleotides. The biosensor enabled distinction between DNA samples isolated from soybean RoundupReady® (RR soybean) and non-genetically modified soybean. The frequent introduction of investigated DNA sequences in other genetically modified organisms (GMOs) give a broad perspectives for analytical application of the biosensor.
机译:提出了一种用于检测转基因食品成分的电化学生物传感器。该生物传感器基于对35S启动子或nos终止子具有特异性的21-mer单链寡核苷酸(ssDNA探针),它们通常存在于转基因DNA盒中。使用活化试剂–水溶性1-乙基-3(3'-二甲基氨基丙基)-碳二亚胺(ssDNA探针)在金电极表面的半胱胺自组装单分子膜(SAM)的氨基末端5'-磷酸末端共价连接EDC)和N-羟基磺基琥珀酰亚胺(NHS)。电极表面的杂交反应是通过亚甲基蓝(MB)检测的,该亚甲基蓝对ssDNA探针的亲和力高于对DNA双链体的亲和力。使用19-mer oligoG和oligoC核苷酸优化了电极修饰程序。生物传感器可以区分从大豆RoundupReady ®(RR大豆)和非转基因大豆中分离的DNA样品。在其他转基因生物(GMO)中频繁引入被研究的DNA序列,为生物传感器的分析应用提供了广阔的前景。

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