title: New qualitative detection methods of genetically modified potatoes.
abstract: In Japan, 8 lines of genetically modified (GM) potato (2 lines of NewLeaf potato; NL, 3 lines of NewLeaf Plus potato; NLP, and 3 lines of NewLeaf Y potato; NLY) have already been authorized as safe for use in foods and feeds. We have developed polymerase chain reaction (PCR) methods for the qualitative detection of the GM potatoes for the screening and the identification of NL, NLP and NLY. The gene encoding uridine diphosphate (UDP)-glucose pyrophosphorylase (UGPase) was used as a taxon specific gene. We designed the primer pair to detect the cryIIIA genes as a screening method for GM potatoes because the gene should be inserted in all 8 lines of the GM potatoes. For identification of NL, NLP and NLY, we further designed three specific primer pairs for the different recombinant DNAs (r-DNA) specifically introduced into NL, NLP, or NLY. In addition, to identify the 3 lines of NLY that have been introduced with the same r-DNA, the three line-specific primer pairs for the border sequence between the r-DNA and genomic DNA of NLY 3 lines were designed. Six lines of GM potato used as the test material were specifically identified using the each primer pair under the same PCR condition. The detection limits of all the GM potatoes should be approximately 0.1%. Furthermore, the specificity and reproducibility of the methods were confirmed in a six-laboratory collaborative study.
authors.affiliation: National Institute of Health Sciences, 1-18-1 Kamiyoga, Setagaya-ku, Tokyo 158-8501, Japan. tawata@nihs.go.jp
authors.forename: Takahiro
authors.initials: T
authors.lastname: Watanabe
authors.forename: Hideo
authors.initials: H
authors.lastname: Kuribara
authors.forename: Takashi
authors.initials: T
authors.lastname: Mishima
authors.forename: Hiroyuki
authors.initials: H
authors.lastname: Kikuchi
authors.forename: Takashi
authors.initials: T
authors.lastname: Kodama
authors.forename: Satoshi
authors.initials: S
authors.lastname: Futo
authors.forename: Kikuko
authors.initials: K
authors.lastname: Kasama
authors.forename: Akie
authors.initials: A
authors.lastname: Toyota
authors.forename: Masanori
authors.initials: M
authors.lastname: Nouno
authors.forename: Ayako
authors.initials: A
authors.lastname: Saita
authors.forename: Kunihiko
authors.initials: K
authors.lastname: Takahashi
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
authors.forename: Misao
authors.initials: M
authors.lastname: Kubo
mesh_terms: D000083722:Bacillus thuringiensis Toxins; D001426:Bacterial Proteins / Q000737:chemistry / Q000235:genetics; D001427:Bacterial Toxins / Q000737:chemistry / Q000235:genetics; D017931:DNA Primers; D004274:DNA, Recombinant / Q000302:isolation & purification; D004731:Endotoxins / Q000737:chemistry / Q000235:genetics; D006460:Hemolysin Proteins; D007564:Japan; D030821:Plants, Genetically Modified / Q000235:genetics*; D016133:Polymerase Chain Reaction / Q000379:methods*; D012680:Sensitivity and Specificity; D011198:Solanum tuberosum / Q000235:genetics*; D005957:UTP-Glucose-1-Phosphate Uridylyltransferase / Q000737:chemistry / Q000235:genetics
chemical_list: D000083722:Bacillus thuringiensis Toxins; D001426:Bacterial Proteins; D001427:Bacterial Toxins; D017931:DNA Primers; D004274:DNA, Recombinant; D004731:Endotoxins; D006460:Hemolysin Proteins; C030956:insecticidal crystal protein, Bacillus Thuringiensis; D005957:UTP-Glucose-1-Phosphate Uridylyltransferase