A Novel Detection System for the Genetically Modified Canola (Brassica rapa) Line RT73
A Novel Detection System for the Genetically Modified Canola (Brassica rapa) Line RT73
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DOI：
10.1021/ac102434q
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发表时间：
2010-12-01
影响因子：
7.4
通讯作者：
Teshima, Reiko
中科院分区：
文献类型：
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作者：
Akiyama, Hiroshi;Makiyama, Daiki;Teshima, Reiko
抗除草剂转基因油菜（Brass:ea napus）系RT73已在全球范围内被批准用于动物饲料和人类食品。然而，与RT73 B油菜种间杂交而成的RT73油菜系尚未在日本获得批准。我们开发了一种双相实时聚合酶链反应（PCR）方法，利用终点分析的散点图来区分油菜B和油菜B的DNA，该方法能够区分油菜B和芥菜组与油菜B、油菜籽B、油菜籽B、油菜籽B和油菜B的DNA。我们还开发了一种双相实时PCR方法，用于同时检测RT73特异性序列和内源FatA基因。此外，我们还开发并优化了96孔二氧化硅膜板dna提取方法，用于单个油菜籽粒样品的dna提取。我们的检测系统可以识别油菜籽粒样品中的RT73 B rapa籽粒，从而准确可靠地监测油菜中RT73 B rapa的污染情况。从而在日本的政府监管中发挥了作用
The herbicide tolerant genetically modified Roundup Ready canola (Brass:ea napus) line RT73 has been approved worldwide for use in animal feed and human food However, RT73 Brassica rapa lines derived from inter specific crosses with RT73 B napus have not been approved in Japan Here, we report on a novel system using individual kernel analyses for the qualitative detection of RT73 B rapa in canola grain samples We developed a duplex real time polymerase chain reaction (PCR) method to discriminate B napus and B rapa DNA using scatter plots of the end point analyses, this method was able to discriminate a group comprising B rapa and Brassica juncea from a group comprising B napus, Brassica carinata, and Brassie-a oleracea We also developed a duplex real-time PCR method for the simultaneous detection of an RT73-specific sequence and an endogenous FatA gene Additionally, a DNA-extraction method using 96 well silica membrane plates was developed and optimized for use with individual canola kernels Our detection system could identify RT73 B rapa kernels in canola grain samples enabling the accurate and reliable monitoring of RT73 B rapa contamination in canola, thus playing a role in its governmental regulation in Japan