title: Real-Time PCR-Based Quantitation Method for the Genetically Modified Soybean Line GTS 40-3-2.
abstract: This chapter describes a real-time PCR-based method for quantitation of the relative amount of genetically modified (GM) soybean line GTS 40-3-2 [Roundup Ready(®) soybean (RRS)] contained in a batch. The method targets a taxon-specific soybean gene (lectin gene, Le1) and the specific DNA construct junction region between the Petunia hybrida chloroplast transit peptide sequence and the Agrobacterium 5-enolpyruvylshikimate-3-phosphate synthase gene (epsps) sequence present in GTS 40-3-2. The method employs plasmid pMulSL2 as a reference material in order to quantify the relative amount of GTS 40-3-2 in soybean samples using a conversion factor (Cf) equal to the ratio of the RRS-specific DNA to the taxon-specific DNA in representative genuine GTS 40-3-2 seeds.
authors.affiliation: National Food Research Institute, National Agriculture and Food Research Organization, 2-1-12 Kannon-dai, Tsukuba, 305-8642, Japan. kaz@affrc.go.jp.
authors.forename: Kazumi
authors.initials: K
authors.lastname: Kitta
authors.affiliation: National Food Research Institute, National Agriculture and Food Research Organization, 2-1-12 Kannon-dai, Tsukuba, 305-8642, Japan.
authors.forename: Reona
authors.initials: R
authors.lastname: Takabatake
authors.affiliation: National Food Research Institute, National Agriculture and Food Research Organization, 2-1-12 Kannon-dai, Tsukuba, 305-8642, Japan.
authors.forename: Junichi
authors.initials: J
authors.lastname: Mano
mesh_terms: D051229:3-Phosphoshikimate 1-Carboxyvinyltransferase; D060054:Agrobacterium / Q000378:metabolism; D001426:Bacterial Proteins; D060365:Chloroplast Proteins; D015870:Gene Expression; D032306:Petunia / Q000378:metabolism; D037121:Plant Lectins / Q000235:genetics*; D030821:Plants, Genetically Modified; D021382:Protein Sorting Signals; D012333:RNA, Messenger; D060888:Real-Time Polymerase Chain Reaction; D011993:Recombinant Fusion Proteins / Q000235:genetics*; D013025:Glycine max / Q000235:genetics* / Q000378:metabolism
chemical_list: D001426:Bacterial Proteins; D060365:Chloroplast Proteins; D037121:Plant Lectins; D021382:Protein Sorting Signals; D012333:RNA, Messenger; D011993:Recombinant Fusion Proteins; C078737:chloroplast transit peptides; D051229:3-Phosphoshikimate 1-Carboxyvinyltransferase
keywords: Conversion factor; GTS 40-3-2; Genetically modified soybeans; Real-time PCR; TaqMan® chemistry