Molecular Detection Of Genetically Modified Soybean Establishment And Application Of Techniacl Methods

With the development of bio-engineering, transgenic technology plays an irreplaceable role in modern agriculture, more and more genetically modified crops were planted worldwide.In this paper, the assessment methods of genetically modified crops were conduct with taking transgenic soybean plants as a example. Soybean is one of the important crops in modern agriculture, not only it is rich in protein, vegetable oil and a variety of ingredients beneficial to human body, but also it is the main source of vegetable protein in human food and important industrial raw materials and livestock feed. Currently, soybean production gradually begins unable to meet the demand of people’s needs. The application of transgenic technology is one of the most effective way to increase soybean yield in China. Since the first successful soybean transformation was reported, many soybean transformation examples were reported one after another. With the vast commercial cultivation of genetically modified crops, the safety concern on genetically modified crops has raised close attention. Therefore, to guarantee the interests of consumers, the assessment of the transgenic plant is obligatory. Only detection of genetically modified crops content and composition were accurately accessed, the monitoring of genetically modified crops can be keeping pace with the times. Work in this paper was as followed:1. The CaMV35promoter, NOS terminator and the NPTII gene elements were accessed and used preliminary to adjust whether the crops is transgenic crops,which lays the foundation for further specific qualitative detection.2. Special primers based on transgenic soybean target gene GmCBL and NAC5genome sequences and CaMV35S promoter were designed. With these two primers, we established a genetically modified soybean GmCBL and NAC5strain-specific qualitative PCR detection method. The PCR products of the detection by this method not only contain the genome sequence, but also contain the target gene sequence.Because of its highly specificity, false-positive results due to Broccoli.Mosaic virus infected was exclude.3. Genetically modified soybeans Gm7a subunit was taken as a example for qualitative detection. The copy number of the target gene Gm7a in transgenic soybean were obtained by using real-time fluorescence quantitative PCR method with Tubulin as a reference gene and a standard curve by using positive plasmid containing the BAR gene.