New qualitative detection methods of genetically modified potatoes.
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.
Watanabe
Takahiro
T
National Institute of Health Sciences, 1-18-1 Kamiyoga, Setagaya-ku, Tokyo 158-8501, Japan. tawata@nihs.go.jp
Kuribara
Hideo
H
Mishima
Takashi
T
Kikuchi
Hiroyuki
H
Kodama
Takashi
T
Futo
Satoshi
S
Kasama
Kikuko
K
Toyota
Akie
A
Nouno
Masanori
M
Saita
Ayako
A
Takahashi
Kunihiko
K
Hino
Akihiro
A
Akiyama
Hiroshi
H
Maitani
Tamio
T
Kubo
Misao
M
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
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