Ministry of Education Key Laboratory of Analysis and Detection Technology for Food Safety Fuzhou University, Department of Chemistry, Fuzhou University, Fuzhou 350002, China.
Biosens Bioelectron. 2013 Mar 15;41:168-71. doi: 10.1016/j.bios.2012.08.017. Epub 2012 Aug 17.
In this paper, we reported a convenient fluorescence method for the detection of genetically modified organisms (GMOs). As it is known that the cauliflower mosaic virus (CaMV) 35S promoter is widely used in most transgenic plants (Schnurr and Guerra, 2000), we thus design a simple method based on the detection of a section target DNA (DNA-T) from the transgene CaMV 35S promoter. In this method, the full-length guanine-rich single-strand sequences were split into fragments (Probe 1 and 2) and each part of the fragment possesses two GGG repeats. In the presence of K(+) ion and berberine, if a complementary target DNA of the CaMV 35S promoter was introduced to hybridize with Probe 1 and 2, a G-quadruplex-berberine complex was thus formed and generated a strong fluorescence signal. The generation of fluorescence signal indicates the presence of CaMV 35S promoter. This method is able to identify and quantify Genetically Modified Organisms (GMOs), and it shows wide linear ranges from 5.0×10(-9) to 9.0×10(-7) mol/L with a detection limit of 2.0×10(-9) mol/L.
本文报道了一种用于检测转基因生物（GMO）的简便荧光方法。众所周知，花椰菜花叶病毒（CaMV）35S 启动子被广泛应用于大多数转基因植物（Schnurr 和 Guerra，2000），因此我们设计了一种基于检测转基因 CaMV 35S 启动子的一段目标 DNA（DNA-T）的简单方法。在该方法中，全长富含鸟嘌呤的单链序列被分成片段（探针 1 和 2），每个片段都含有两个 GGG 重复序列。在 K+离子和小檗碱存在的情况下，如果引入与 CaMV 35S 启动子互补的目标 DNA 与探针 1 和 2杂交，就会形成 G-四链体-小檗碱复合物，从而产生强烈的荧光信号。荧光信号的产生表明 CaMV 35S 启动子的存在。该方法能够识别和定量转基因生物（GMO），具有较宽的线性范围，从 5.0×10(-9)到 9.0×10(-7)mol/L，检测限为 2.0×10(-9)mol/L。
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