Development of a Screening Method for Genetically Modified Soybean by Plasmid-Based Quantitative Competitive Polymerase Chain Reaction
2008
Shimizu, E. | Kato, H. | Nakagawa, Y. | Kodama, T. | Futo, S. | Minegishi, Y. | Watanabe, T. | Akiyama, H. | Teshima, R. | Furui, S.
A novel type of quantitative competitive polymerase chain reaction (QC-PCR) system for the detection and quantification of the Roundup Ready soybean (RRS) was developed. This system was designed based on the advantage of a fully validated real-time PCR method used for the quantification of RRS in Japan. A plasmid was constructed as a competitor plasmid for the detection and quantification of genetically modified soy, RRS. The plasmid contained the construct-specific sequence of RRS and the taxon-specific sequence of lectin1 (Le1), and both had 21 bp oligonucleotide insertion in the sequences. The plasmid DNA was used as a reference molecule instead of ground seeds, which enabled us to precisely and stably adjust the copy number of targets. The present study demonstrated that the novel plasmid-based QC-PCR method could be a simple and feasible alternative to the real-time PCR method used for the quantification of genetically modified organism contents.
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