A Multiplex PCR Method of Detecting Recombinant DNAs from Five Lines of Genetically Modified Maize.

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  • MATSUOKA Takeshi
    National Food Research Institute, Ministry of Agriculture, Forestry and Fisheries Tokyo Center for Quality Control and Consumer Service, Ministry of Agriculture, Forestry and Fisheries
  • KURIBARA Hideo
    National Food Research Institute, Ministry of Agriculture, Forestry and Fisheries Tokyo Center for Quality Control and Consumer Service, Ministry of Agriculture, Forestry and Fisheries
  • AKIYAMA Hiroshi
    National Institute of Health Sciences
  • MIURA Hirohito
    National Food Research Institute, Ministry of Agriculture, Forestry and Fisheries
  • GODA Yukihiro
    National Institute of Health Sciences
  • KUSAKABE Yuko
    National Food Research Institute, Ministry of Agriculture, Forestry and Fisheries
  • ISSHIKI Kenji
    National Food Research Institute, Ministry of Agriculture, Forestry and Fisheries
  • TOYODA Masatake
    National Institute of Health Sciences
  • HINO Akihiro
    National Food Research Institute, Ministry of Agriculture, Forestry and Fisheries

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Other Title
  • 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
  • Multiplex PCR Method of Detecting Recombinant DNAs from Five Lines of Genetically Modified Maize

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Abstract

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.

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