Real-time loop-mediated isothermal amplification for the CaMV-35S promoter as a screening method for genetically modified organisms
Real-time loop-mediated isothermal amplification for the CaMV-35S promoter as a screening method for genetically modified organisms
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DOI：
10.1007/s00217-003-0862-5
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发表时间：
2004-04-01
影响因子：
3.3
通讯作者：
Kanbe, M
中科院分区：
文献类型：
--
作者：
Fukuta, S;Mizukami, Y;Kanbe, M
已经开发了一种检测转基因生物的方法。检测方法基于环介导等温扩增（LAMP）反应。利用一对LAMP引物扩增了花椰菜花叶病毒35 S（CaMV-35 S）的启动子基因。LAMP方法的特征之一是其能够合成极大量的DNA。因此，产生大量副产物焦磷酸根离子，在LAMP反应混合物中产生焦磷酸镁的白色沉淀。通过测定反应液的浊度，不需要凝胶电泳，就可以容易地检测CaMV-35 S启动子基因的扩增。利用这项新技术，我们分析了抗农达大豆，发现转基因含量从0.5%到5%不等。
A method has been developed to detect genetically modified organisms (GMOs). The detection method is based on the loop-mediated isothermal amplification (LAMP) reaction. Cauliflower mosaic virus 35S (CaMV-35S) promoter gene, a widespread genetic element, was amplified by a set of LAMP primers. One of the characteristics of the LAMP method is its ability to synthesize an extremely large amount of DNA. Accordingly, a large amount of byproduct, pyrophosphate ion, is produced, yielding a white precipitate of magnesium pyrophosphate in the LAMP reaction mixture. The measurement of the turbidity of the reaction mixture allows easy detection of amplification of CaMV-35S promoter gene without the need for gel electrophoresis. With this new technique we analyzed Roundup Ready soybean, and found GMO content ranging from 0.5% to 5%.