An Endogenous Reference Gene of Common and Durum Wheat for Detection of Genetically Modified Wheat

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  • IMAI Shinjiro
    Quality Assurance Division, Research Center for Basic Science Research and Development, Nisshin Seifun Group Inc.
  • TANAKA Keiko
    Quality Assurance Division, Research Center for Basic Science Research and Development, Nisshin Seifun Group Inc.
  • NISHITSUJI Yasuyuki
    Quality Assurance Division, Research Center for Basic Science Research and Development, Nisshin Seifun Group Inc.
  • KIKUCHI Yosuke
    Quality Assurance Division, Research Center for Basic Science Research and Development, Nisshin Seifun Group Inc.
  • MATSUOKA Yasuyuki
    Central Laboratory, Nippon Flour Mills Co., Ltd.
  • ARAMI Shin-ichiro
    Central Laboratory, Nippon Flour Mills Co., Ltd.
  • SATO Megumi
    Central Laboratory, Nippon Flour Mills Co., Ltd.
  • HARAGUCHI Hiroyuki
    Central Laboratory, Nippon Flour Mills Co., Ltd.
  • KURIMOTO Youichi
    Central Laboratory, Nippon Flour Mills Co., Ltd.
  • MANO Junichi
    National Agriculture and Food Research Organization, National Food Research Institute
  • FURUI Satoshi
    National Agriculture and Food Research Organization, National Food Research Institute
  • KITTA Kazumi
    National Agriculture and Food Research Organization, National Food Research Institute

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  • 遺伝子組換えコムギ検知に有用な普通コムギおよびデュラムコムギの内在性遺伝子

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Abstract

To develop a method for detecting GM wheat that may be marketed in the near future, we evaluated the proline-rich protein (PRP) gene as an endogenous reference gene of common wheat (Triticum aestivum L.) and durum wheat (Triticum durum L.). Real-time PCR analysis showed that only DNA of wheat was amplified and no amplification product was observed for phylogenetically related cereals, indicating that the PRP detection system is specific to wheat. The intensities of the amplification products and Ct values among all wheat samples used in this study were very similar, with no nonspecific or additional amplification, indicating that the PRP detection system has high sequence stability. The limit of detection was estimated at 5 haploid genome copies. The PRP region was demonstrated to be present as a single or double copy in the common wheat haploid genome. Furthermore, the PRP detection system showed a highly linear relationship between Ct values and the amount of plasmid DNA, indicating that an appropriate calibration curve could be constructed for quantitative detection of GM wheat. All these results indicate that the PRP gene is a suitable endogenous reference gene for PCR-based detection of GM wheat.

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