A method of detecting recombinant DNAs from four lines of genetically modified maize.
A method of detecting recombinant DNAs from four lines of genetically modified maize.
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一种检测来自四个转基因玉米品系的重组 DNA 的方法。
DOI：
10.3358/shokueishi.41.137
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发表时间：
2000
期刊：
影响因子：
--
通讯作者：
A. Hino
中科院分区：
文献类型：
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作者：
T. Matsuoka;Y. Kawashima;H. Akiyama;H. Miura;Y. Goda;Y. Kusakabe;K. Isshiki;M. Toyoda;A. Hino
建立了转基因玉米（GM-玉米）的PCR检测方法。从美国引进的4个转基因玉米品系中，根据新导入基因的序列设计了4对特异性寡核苷酸引物，可以检测到转基因玉米中重组脱氧核糖核酸（DNA）的存在。还检测玉米醇溶蛋白基因作为内部对照。该方法允许通过使用特异性引物对特异性检测Bt 11、Event 176、MON 810和LIBERTY中的每一个，所述特异性引物对被设计用于扩增包括部分外源引入的序列和部分内在玉米序列的区段。Event 176、MON 810和LIBERTY的检测灵敏度约为0.05%，Bt 11的检测灵敏度约为0.01%。为了区分三个抗虫转基因玉米品系，我们设计了一种多重PCR方法。这三个GM-玉米系基于其扩增子的预期长度是可区分的。
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.