Development of a screening method for genetically modified soybean by plasmid-based quantitative competitive polymerase chain reaction.
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.
Shimizu
Eri
E
Fasmac Co., Ltd., 5-1-3 Midorigaoka, Atsugi, Kanagawa 243-0041, Japan.
Kato
Hisashi
H
Nakagawa
Yuki
Y
Kodama
Takashi
T
Futo
Satoshi
S
Minegishi
Yasutaka
Y
Watanabe
Takahiro
T
Akiyama
Hiroshi
H
Teshima
Reiko
R
Furui
Satoshi
S
Hino
Akihiro
A
Kitta
Kazumi
K
D018744:DNA, Plant / Q000032:analysis*; D005998:Glycine / Q000031:analogs & derivatives; D006540:Herbicides; D037121:Plant Lectins / Q000235:genetics; D030821:Plants, Genetically Modified / Q000235:genetics*; D010957:Plasmids / Q000235:genetics*; D016133:Polymerase Chain Reaction / Q000379:methods*; D012639:Seeds / Q000235:genetics; D030262:Soybean Proteins / Q000235:genetics; D013025:Glycine max / Q000145:classification / Q000235:genetics*; D000097797:Glyphosate; D010835:Phytohemagglutinins
D018744:DNA, Plant; D005998:Glycine; D006540:Herbicides; D037121:Plant Lectins; D030262:Soybean Proteins; D000097797:Glyphosate; C028824:soybean lectin; D010835:Phytohemagglutinins