Establishment And Application Of RT-PCR Detection On Glyphosate Resistant Transgenic Soybean In Daily Soy

This paper takes soybean raw material and products as the objects to establish the Realtime-PCR method on the detection of transgenic glyphosate-resistant soybeans(EPSPS). And this method was used to detect the EPSPS the common soybean products in the market. The establishment of this method can provide the theoretical and experimental basis and reference for the food regulatory authorities to effective management in genetically modified products.Regarding the influence of PVP concentration on the extraction of soybean genomic DNA, soybean samples from three districts were analyzed. PCR reaction and native gel electrophoresis analysis were conducted to the house-keeping gene Lectin. The result shown that the concentration of PVP was over 0.1% and can effectively improve the extraction quality of soybean genomic DNA. The location of the target gene can be obviously seen in the gel electrophoresis analysis and the bands.In the establishment on the detection of the transgenic glyphosate-resistant soybeans in soybean sauce made by transgenic soybean residue, the fluorescence PCR reaction(Syb Green) was used to detect the genomic DNA extracted from soybean sauce. The Ca Mv35 s promoter and Nos terminator regulatory elements were detected in the genomic DNA extracted from soybean sauce. Meanwhile, the degradation of exogenous gene EPSPS gene was relatively serious. It can be confirmed that the two regulatory elements Ca Mv35 s promoter and Nos terminator can be used as the detection targets to distinguish whether the soybean sauce is produced by the soybean residue of transgenic glyphosate-resistant soybeans. Compared to the traditional PCR, Realtime fluorescence quantitative PCR was a great potential method in the detection of genetically modified food with the advantages of faster, higher throughput, higher selectivity and higher sensitivity. In the detection of transgenic glyphosate-resistant soybeans in common soybean products in the market, 28 kinds of soybean products are taken as sampling including the soybean products in bulk from agricultural markets of and those in bags from supermarkets. Fluorescence PCR was used to detect the transgenic glyphosateresistant soybeans. In eleven kinds of dried bean curd products in bags, two kinds of them contained Ca Mv35 s promoter, Nos terminator and EPSPS gene of transgenic glyphosate-resistant soybeans. One kind of them contained Ca Mv35 s promoter and Nos terminator of transgenic glyphosate-resistant soybeans. In 17 kinds of soybean products in bulk, only one kind of them i contained Ca Mv35 s promoter, Nos terminator and EPSPS gene. In 20 samples, no component of transgenic glyphosate-resistant soybeans was detected in the products whose labels clearly label that the raw material is nonGMO soybean. As for the products which don't label whether it contains transgenic soybean material, four of them are detected to contain the transgenic glyphosateresistant soybeans.