title: Detection method for genetically modified papaya using duplex PCR.
abstract: A simple and rapid method for the identification of genetically modified (GM) papaya, derived from Line 55-1, was developed by modifying the Japanese official PCR method. Genomic DNA was directly extracted from the fresh fruit without the lyophilization step, using a commercial silica-based kit. To develop a duplex PCR method which simultaneously detects the GM papaya-specific gene and the intrinsic papain gene, the papain 2-5'/3' (amplicon size; 184 bp) primer pair for the detection of the papain gene was newly designed within the region of the products (211 bp) amplified using the papain 1-5'/-3' primer pair adopted in the Japanese official PCR method. To detect the GM papaya-specific gene, the primer pair Nos C-5'/CaM N-3' described in the Japanese official method was used. The DNA sequences of the GM papaya gene and the intrinsic papain gene were co-amplified using the PCR method in a single tube. The developed duplex PCR method allows the simultaneous detection of the products by means of agarose gel electrophoresis or microchip electrophoresis. The proposed method for GM papaya identification is simple and rapid.
authors.affiliation: Japan Food Research Laboratories, Chitose: 2-3, Bunkyo, Chitose-shi, Hokkaido 066-0052, Japan.
authors.forename: Akihiro
authors.initials: A
authors.lastname: Yamaguchi
authors.forename: Kaori
authors.initials: K
authors.lastname: Shimizu
authors.forename: Takashi
authors.initials: T
authors.lastname: Mishima
authors.forename: Nobutaro
authors.initials: N
authors.lastname: Aoki
authors.forename: Hideki
authors.initials: H
authors.lastname: Hattori
authors.forename: Hidetaka
authors.initials: H
authors.lastname: Sato
authors.forename: Nobuo
authors.initials: N
authors.lastname: Ueda
authors.forename: Takahiro
authors.initials: T
authors.lastname: Watanabe
authors.forename: Akihiro
authors.initials: A
authors.lastname: Hino
authors.forename: Hiroshi
authors.initials: H
authors.lastname: Akiyama
authors.forename: Tamio
authors.initials: T
authors.lastname: Maitani
mesh_terms: D029441:Carica / Q000235:genetics*; D018744:DNA, Plant / Q000302:isolation & purification; D005513:Food Inspection / Q000379:methods*; D030841:Food, Genetically Modified; D018745:Genome, Plant / Q000235:genetics; D016133:Polymerase Chain Reaction / Q000379:methods*
chemical_list: D018744:DNA, Plant