Multiplex PCR法を用いた組換えトウモロコシ5系統からの組換え遺伝子の検知法

DOI 日本農学文献記事索引 Web Site PubMed 被引用文献20件 参考文献18件

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タイトル別名
  • A Multiplex PCR Method of Detecting Recombinant DNAs from Five Lines of Genetically Modified Maize.
  • Multiplex PCR Method of Detecting Recombinant DNAs from Five Lines of Genetically Modified Maize

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Seven lines of genetically modified (GM) maize have been authorized in Japan as foods and feeds imported from the USA. We improved a multiplex PCR method described in the previous report in order to distinguish the five lines of GM maize. Genomic DNA was extracted from GM maize with a silica spin column kit, which could reduce experimental time and improve safety in the laboratory and potentially in the environment. We sequenced recombinant DNA (r-DNA) introduced into GM maize, and re-designed new primer pairs to increase the specificity of PCR to distinguish five lines of GM maize by multiplex PCR. A primer pair for the maize intrinsic zein gene (Ze1) was also designed to confirm the presence of amplifiable maize DNA. The lengths of PCR products using these six primer pairs were different. The Ze1 and the r-DNAs from the five lines of GM maize were qualitatively detected in one tube. The specific PCR bands were distinguishable from each other on the basis of the expected length. The r-DNA could be detected from maize samples containing 0.5% of each of the five lines of GM maize. The sensitivity would be acceptable to secure the verification of non-GMO materials and to monitor the reliability of the labeling system.

収録刊行物

  • 食品衛生学雑誌

    食品衛生学雑誌 42 (1), 24-32, 2001

    公益社団法人 日本食品衛生学会

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