A method of detecting recombinant DNAs from four lines of genetically modified maize.

T. Matsuoka, Y. Kawashima, H. Akiyama, H. Miura, Y. Goda, Y. Kusakabe, K. Isshiki, M. Toyoda, A. Hino
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引用次数: 84

Abstract

A method using polymerase chain reaction (PCR) was designed for the detection of genetically modified maize (GM-maize). There are four lines of GM-maize imported from the United States, and the presence of recombinant deoxyribonucleic acid (DNA) in the maize could be detected with four pairs of specific oligonucleotide primers designed from the sequences of the newly introduced genes. The maize zein gene was also detected as an internal control. This method allows specific detection of each of Bt11, Event176, MON810 and LIBERTY by using pairs of specific primers designed to amplify a segment including part of the exogenously introduced sequence and part of the intrinsic maize sequence. The detection sensitivity was about 0.05% for Event176, MON810 and LIBERTY, and about 0.01% for Bt11. To distinguish among three insect-resistant GM-maize lines, we designed a multiplex PCR method. These three GM-maize lines were distinguishable on the basis of the expected lengths of their amplicons.
一种检测四种转基因玉米系重组dna的方法。
设计了一种检测转基因玉米(GM-maize)的聚合酶链反应(PCR)方法。从美国引进的转基因玉米有4个品系,利用新引进基因序列设计的4对特异性寡核苷酸引物可检测玉米中重组脱氧核糖核酸(DNA)的存在。玉米蛋白基因也被检测为内控基因。该方法通过使用特定的引物对Bt11、Event176、MON810和LIBERTY进行特异性检测,该引物设计用于扩增片段,其中包括部分外源引入序列和部分内在玉米序列。对Event176、MON810和LIBERTY的检测灵敏度约为0.05%,对Bt11的检测灵敏度约为0.01%。为了区分3个抗虫转基因玉米品系,我们设计了多重PCR方法。这三种转基因玉米系是根据其扩增子的预期长度来区分的。
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