title: An endogenous reference gene of common and durum wheat for detection of genetically modified wheat.
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.
authors.affiliation: Quality Assurance Division, Research Center for Basic Science Research and Development, Nisshin Seifun Group Inc., Saitama, Japan.
authors.forename: Shinjiro
authors.initials: S
authors.lastname: Imai
authors.forename: Keiko
authors.initials: K
authors.lastname: Tanaka
authors.forename: Yasuyuki
authors.initials: Y
authors.lastname: Nishitsuji
authors.forename: Yosuke
authors.initials: Y
authors.lastname: Kikuchi
authors.forename: Yasuyuki
authors.initials: Y
authors.lastname: Matsuoka
authors.forename: Shin-Ichiro
authors.initials: S
authors.lastname: Arami
authors.forename: Megumi
authors.initials: M
authors.lastname: Sato
authors.forename: Hiroyuki
authors.initials: H
authors.lastname: Haraguchi
authors.forename: Youichi
authors.initials: Y
authors.lastname: Kurimoto
authors.forename: Junichi
authors.initials: J
authors.lastname: Mano
authors.forename: Satoshi
authors.initials: S
authors.lastname: Furui
authors.forename: Kazumi
authors.initials: K
authors.lastname: Kitta
mesh_terms: D018744:DNA, Plant / Q000032:analysis; D030841:Food, Genetically Modified; D010940:Plant Proteins / Q000235:genetics; D060888:Real-Time Polymerase Chain Reaction; D014908:Triticum / Q000235:genetics*
chemical_list: D018744:DNA, Plant; D010940:Plant Proteins; C070314:WPRP1 protein, wheat