Development of a multiplex and semi-quantitative PCR assay for detection of genetically modified Roundup Ready Soybean in some representative food

• Main Authors: Wei-Yi Wang, 王薇猗
• Other Authors: Tony J. Fang
• Format: Others
• Language: zh-TW
• Published: 2002
• Online Access: http://ndltd.ncl.edu.tw/handle/78913409446366468242

碩士 === 國立中興大學 === 食品科學系 === 90 === Abstract Genetically modified organisms (GMO) have raised much attention in recent years. Uptoday, great deals of genetically modified organisms are sold in the market. Over 50% of imported soybeans, mainly Roundup ReadyTM soybean, in Taiwan were genetically modified. Department of Health (DOH), The Executive Yuan, Republic of China has published “Safety assessment methodology for Genetically modified foods (GMF)” and “Labeling act for Genetically modified foods (GMF)”, which is the foundation of management of GMF in Taiwan. This studied was to develope the qualitative and semi-quantitative detection method for genetically modified soybean by using the multiplex PCR technique. The potential application of the developed methods in detection of GM soybeans raw material or products were also evaluated. In this stduy, various primers for detection of cloned genes in Roundup ReadyTM, namely, 35SP-F/R (143 bp), NOS-F/R (189 bp), EPSPS-F/R (314 bp) and 35SP-F/CTP-R (256 bp), were artificially synthezied. In addtion, primer LEC-F/R (175 bp) was used for detection of species specificity of soybeans. The results showed that the samples containing genetically modified soybeans were detected by multiplex PCR with primers 35SP-F/CTP-R and LEC-F/R. Semi-quantitative analysis of PCR products was developed by using Gel-Pro Analyzer and Sigma Plot softwere with standard curves of DNA that contained various ratios of GMO. This method was used to estimate the amount of GM soybeans in retail products that was made of soybeans. Our results showed that GM soybeans were not only easily detected by multiplex PCR and semi-quantitative method the detection time and cost could also be reduced significantly.