We analyzed the DNA fragments extracted from four rice vermicelli products. The Bacillus thuringiensis (Bt) rice line, which has a construct similar to the GM Shanyou 63 line, was detected in some vermicelli products by identification of the junction region sequence between rice Act1 promoter and the Cry1Ac gene, and that between Cry1Ac and nos. In addition, we also detected a different Bt rice line by means of the junction region sequence between the maize ubiquitin promoter and cry1Ab gene and that between the cauliflower mosaic virus 35S promoter and the hygromycin phosphotransferase in some vermicelli products. Accordingly, we for the first time have detected the two transgenic Bt rice lines contaminating rice vermicelli samples. Furthermore, we developed a duplex real-time polymerase chain reaction (PCR) method for the simultaneous detection of both Bt rice lines.
Many kinds of genetically modified (GM) crops, which include GM soy, maize, rapeseed, cotton and potato, have already been developed and the cultivated acreage of these crops has continued to grow year by year. It was reported that the global area of GM crops for 2003 was 67.7 million hectares with a growth rate of 15% compared to that in 2002. This growth is estimated to rapidly increase, since the planting has been spread all over the world in addition to the nations such as United States (U.S.) and Canada. 1) On the other hand, public concern has been raised in terms of food safety and environmental effects of the GM crops. Especially, consumers are concerned about the negative effects of GM food on their health by their consumption and scientific information has been strongly required.2) Therefore, many governments have now been considering regulations for the use and implementing a labeling system for GM crops as food and feed. Thus, new labeling systems have been introduced for GM foods in the European Union (EU), Australia, Korea, Japan and other countries.The commercialization of fifty-five lines of safety-assessed GM crops including soy, maize, potato, rapeseed, cotton and sugar beet, have already been approved by the Ministry of Health, Labour and Welfare (HMLW) in Japan. To monitor the labeling system, it is necessary to develop reliable and practical methods for the detection and identification of GM crops. 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. 3-10)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. 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 b...
Because the labeling of grains and feed- and foodstuffs is mandatory if the genetically modified organism (GMO) content exceeds a certain level of approved genetically modified varieties in many countries, there is a need for a rapid and useful method of GMO quantification in food samples. In this study, a rapid detection system was developed for Roundup Ready Soybean (RRS) quantification using a combination of a capillary-type real-time PCR system, a LightCycler real-time PCR system, and plasmid DNA as the reference standard. In addition, we showed for the first time that the plasmid and genomic DNA should be similar in the established detection system because the PCR efficiencies of using plasmid DNA and using genomic DNA were not significantly different. The conversion factor (Cf) to calculate RRS content (%) was further determined from the average value analyzed in three laboratories. The accuracy and reproducibility of this system for RRS quantification at a level of 5.0% were within a range from 4.46 to 5.07% for RRS content and within a range from 2.0% to 7.0% for the relative standard deviation (RSD) value, respectively. This system rapidly monitored the labeling system and had allowable levels of accuracy and precision.
A method for the rapid determination of 11 medical components found in health foods for weight loss using liquid chromatography/tandem mass spectrometry (LC/MS/MS) has been developed. HPLC separation is performed on an ODS column with the gradient elution method. The mobile phase consists of two solvents. Solvent A is water and solvent B is methanol/acetonitrile (1 : 1), and both contain 0.1% formic acid and 5 mmol/l of ammonium acetate. Each medical component is analyzed with multiple-reaction monitoring (MRM) in both negative and positive modes through electrospray ionization (ESI). The recovery rates of the 11 medical components added to commercially available health foods were 46.3 114% and each coe‹cient of variation was 13.7% or less. It was conˆrmed that this method is applicable to the urgent analysis of health foods that have caused damage to health.
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