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Electrochemiluminescence-PCR detection of genetically modified organisms

机译:电化学发光-PCR检测转基因生物

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摘要

The detection methods for genetically modified (GM) components in foods have been developed recently. But many of them are complicated and time-consuming; some of them need to use the carcinogenic substance, and can't avoid false-positive results. In this study, an electrochemiluminescence polymerase chain reaction (ECL-PCR) method for detection GM tobaccos is proposed. The Cauliflower mosaic virus 35S (CaMV35S) promoter was amplified by PCR, Then hybridized with a Ru(bpy)32+ (TBR)-labeled and a biotinylated probe. The hybridization products were captured onto streptavidin-coated paramagnetic beads, and detected by measuring the electrochemiluminescence (ECL) signal of the TBR label. Whether the tobaccos contain GM components was discriminated by detecting the ECL signal of CaMV35S promoter. The experiment results show that the detection limit for CaMV35S promoter is 100 fmol, and the GM components can be clearly identified in GM tobaccos. The ECL-PCR method provide a new means in GMOs detection due to its safety, simplicity and high efficiency.
机译:食品中转基因成分的检测方法最近得到了发展。但是其中许多都是复杂且耗时的。其中一些需要使用致癌物质,无法避免假阳性结果。在这项研究中,提出了一种用于检测转基因烟草的电化学发光聚合酶链反应(ECL-PCR)方法。花椰菜花叶病毒35S(CaMV35S)启动子通过PCR扩增,然后与Ru(bpy)32+(TBR)标记的生物素化探针杂交。将杂交产物捕获到涂有链霉亲和素的顺磁珠上,并通过测量TBR标记的电化学发光(ECL)信号进行检测。通过检测CaMV35S启动子的ECL信号来区分烟草是否含有GM成分。实验结果表明,CaMV35S启动子的检出限为100 fmol,在转基因烟草中可以清楚地鉴定出GM成分。 ECL-PCR方法安全,简便,高效,为转基因生物的检测提供了新的手段。

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