Key Laboratory of Plant Resources, Institute of Botany, Chinese Academy of Sciences , Beijing 100093, People's Republic of China.
J Agric Food Chem. 2013 Mar 27;61(12):2964-9. doi: 10.1021/jf3050439. Epub 2013 Mar 15.
The detection and identification of genetically modified (GM) plants are challenging issues that have arisen from the potential negative impacts of extensive cultivation of transgenic plants. The screening process is a long-term focus and needs specific detection strategies. Surface plasmon resonance (SPR) has been used to detect a variety of biomolecules including proteins and nucleic acids due to its ability to monitor specific intermolecular interactions. In the present study, two high-throughput, label-free, and specific methods based on SPR technology were developed to detect transgenic Cry1Ac cotton ( Gossypium spp.) by separately targeting protein and DNA. In the protein-based detection system, monoclonal anti-Cry1Ac antibodies were immobilized on the surface of a CM5 sensor chip. Conventional cotton samples were used to define the detection threshold. Transgenic cotton was easily identified within 5 min per sample. For the DNA-based model, a 25-mer biotinylated oligonucleotide probe was immobilized on an SA sensor chip. PCR products of Cry1Ac (230 bp) were used to investigate the reaction conditions. The sensitivity of the constructed sensor chip was identified at concentrations as low as 0.1 nM based on its complementary base pairing.
检测和鉴定基因修饰(GM)植物是一个具有挑战性的问题,这是由于广泛种植转基因植物可能带来的负面影响所产生的。筛选过程是一个长期的焦点,需要特定的检测策略。由于能够监测特定的分子间相互作用,表面等离子体共振(SPR)已被用于检测各种生物分子,包括蛋白质和核酸。在本研究中,开发了两种基于 SPR 技术的高通量、无标记和特异性方法,分别针对蛋白质和 DNA 来检测转基因 Cry1Ac 棉花(棉属)。在基于蛋白质的检测系统中,将单克隆抗 Cry1Ac 抗体固定在 CM5 传感器芯片的表面上。常规棉花样品用于定义检测阈值。每个样品在 5 分钟内即可轻松识别转基因棉花。对于基于 DNA 的模型,将 25 个碱基的生物素化寡核苷酸探针固定在 SA 传感器芯片上。使用 Cry1Ac 的 PCR 产物(230 bp)来研究反应条件。基于其互补碱基配对,构建的传感器芯片的灵敏度可低至 0.1 nM。