A Detection Method of Recombinant DNA from Genetically Modified Potato(NewLeaf Plus Potato) and Detection of New Leaf Plus Potato in Snack.

Abstract

A detection method using polymerase chain reaction (PCR) was developed to detect the genetically modified (GM) potato (NewLeaf Plus ῌ potato; NL-P), which has not been authorized as safe in foods in Japan. The potato sucrose synthase gene was used as an internal control. The DNA from NL-P specifically provided an amplified band using PCR with a primer pair recognizing PLRV-rep gene. In addition, to prevent false-positive results in processed potato foods infected with PLRV, we designed a primer pair recognizing sequences derived from two organisms to detect specifically NL-P in processed potato. The PCR product obtained using the designed primer pair was specific for NL-P. The DNA introduced into NL-P could be detected from potato powder samples containing 0.05 NL-P. The proposed method was applied to the detection of NL-P in 25 processed potato foods. NL-P was detected in 3 snack products.

New Qualitative Detection Methods of Genetically Modified Potatoes

“…AF078810 (e-p-35S), and the primer pair generated an amplified fragment of 113 bp. In previous studies, 9) p-FMV02-5Ј/PLRV 01-3Ј had been designed in the junction between e-p-35S and cryIIIA for the construct-specific detection of NLP and generated a 234 bp amplification fragment. Moreover, p-FMV05-5Ј/PVY 02-3Ј had been designed in the junction between the 35S promoter sequence of the figwort mosaic virus (P-FMV) and PVYcp for the construct-specific detection of NLY and generated a 225 bp amplified fragment.…”

“…5,9) The amplification products (7.5 ml) of each specific PCR were submitted for electrophoresis at a constant voltage (100 V) on a 3% TAKARA L03 agarose gel (TAKARA Co., Ltd., Tokyo, Japan) in the TAE (40 mmol/l Tris-HCl, 40 mmol/l acetic acid, and 1 mmol/l EDTA, pH 8.0) buffer solution. After electrophoresis, the gel was stained in DW containing 0.5 mg/ml ethidium bromide for 30 min and then washed in DW for 30 min.…”

“…The detection methods based on PCR for GM potatoes have been reported. 9,10,12) To verify the labeling and June 15, 2004 identify of the lines, however, further reliable and practicable detection methods for the GM potatoes are required.…”

“…The polymerase chain reaction (PCR) is one of the widely used systems for the quantitative or qualitative detection of GM crops and we also have previously reported PCR methods for the detection of GM soy, maize, papaya and potatoes. [3][4][5][6][7][8][9][10] The tetraploid cultivated potato (Solanum tuberosum) is one of the world's four major crops and an important feedstuff, but it is easily infected by many kinds of pests and pathogens. 11) Therefore, molecular biology techniques has been attempted to improve the potato varieties which ended with the breeding of GM potatoes commercialized by the Monsanto Co. (St. Louis, MO, U.S.A.).…”

Rapid Quantification Methods for Genetically Modified Maize Contents Using Genomic DNAs Pretreated by Sonication and Restriction Endonuclease Digestion for a Capillary-Type Real-Time PCR System with a Plasmid Reference Standard

“…[2][3][4] The enforcement of these threshold values has created a demand for the development of reliable GMO analysis methods with rapid and inexpensive features. [5][6][7][8][9][10][11][12][13][14][15][16][17][18][19][20][21] Hence many real-time PCR systems based on fluorescent detection, such as TaqMan Ò chemistry, have been developed to identify and quantify GM maize, GM soybeans, and GM varieties of other agricultural commodities. [22][23][24][25][26][27][28][29][30][31][32][33] Real-time PCR systems using TaqMan Ò chemistry are based on the use of a fluorescent TaqMan Ò probe that monitors the formation of the PCR product during each cycle of the reaction.…”