A Screening Method for the Detection of the 35S Promoter and the Nopaline Synthase Terminator in Genetically Modified Organisms in a Real-Time Multiplex Polymerase Chain Reaction Using High-Resolution Melting-Curve Analysis
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- Akiyama Hiroshi
- National Institute of Health Sciences
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- Nakamura Fumi
- National Institute of Health Sciences
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- Yamada Chihiro
- Department of Food Science & Nutrition, Kyoritsu Women's University
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- Nakamura Kosuke
- National Institute of Health Sciences
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- Nakajima Osamu
- National Institute of Health Sciences
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- Kawakami Hiroshi
- Department of Food Science & Nutrition, Kyoritsu Women's University
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- Harikai Naoki
- School of Pharmacy and Pharmaceutical Sciences, Mukogawa Women's University
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- Furui Satoshi
- National Food Research Institute
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- Kitta Kazumi
- National Food Research Institute
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- Teshima Reiko
- National Institute of Health Sciences
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説明
To screen for unauthorized genetically modified organisms (GMO) in the various crops, we developed a multiplex real-time polymerase chain reaction high-resolution melting-curve analysis method for the simultaneous qualitative detection of 35S promoter sequence of cauliflower mosaic virus (35SP) and the nopaline synthase terminator (NOST) in several crops. We selected suitable primer sets for the simultaneous detection of 35SP and NOST and designed the primer set for the detection of spiked ColE1 plasmid to evaluate the validity of the polymerase chain reaction (PCR) analyses. In addition, we optimized the multiplex PCR conditions using the designed primer sets and EvaGreen® as an intercalating dye. The contamination of unauthorized GMO with single copy similar to NK603 maize can be detected as low as 0.1% in a maize sample. Furthermore, we showed that the present method would be applicable in identifying GMO in various crops and foods like authorized GM soybean, authorized GM potato, the biscuit which is contaminated with GM soybeans and the rice which is contaminated with unauthorized GM rice. We consider this method to be a simple and reliable assay for screening for unauthorized GMO in crops and the processing food products.
収録刊行物
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- Biological & Pharmaceutical Bulletin
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Biological & Pharmaceutical Bulletin 32 (11), 1824-1829, 2009
公益社団法人 日本薬学会
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詳細情報 詳細情報について
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- CRID
- 1390282679601194752
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- NII論文ID
- 130000116951
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- NII書誌ID
- AA10885497
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- ISSN
- 13475215
- 09186158
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- NDL書誌ID
- 10412737
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- 本文言語コード
- en
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- データソース種別
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- JaLC
- NDL
- Crossref
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