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转基因大豆305423、MON89788、CV127、GTS40-3-2、356043作为商品化应用最为广泛的大豆品系,种植面积占世界转基因大豆总种植面积的80%以上。根据大豆内标准基因Lectin和5种转基因大豆品系的边界序列设计特异性引物。通过验证引物的适用性、特异性和灵敏度,优化多重PCR检测体系中不同引物的用量及反应退火温度,建立了能同时扩增大豆内源基因Lectin和5个转基因大豆品系的六重PCR检测体系。结果表明:确定的大豆内源基因和5个转基因大豆品系的引物具有很好的特异性,引物之间无交叉扩增和非特异性扩增,多重PCR方法的检测灵敏度达到0.1%。该方法可作为转基因大豆及其产品成分检测的辅助手段,快速检测转基因大豆及其产品中的相应品系。
Genetically modified soybeans 305423, MON89788, CV127, GTS40-3-2, 356043 are the most widely used soybean varieties in commercialization, accounting for more than 80% of the total planting area of the world’s genetically modified soybeans. Specific primers were designed according to the boundary sequences of the standard soybean gene Lectin and five kinds of transgenic soybean lines. Through the verification of the applicability, specificity and sensitivity of the primers, the optimization of the amount of different primers in the multiplex PCR detection system and the reaction annealing temperature, a six-fold PCR detection system that can simultaneously amplify soybean endogenous genes Lectin and five transgenic soybean lines was established . The results showed that the primers of the identified soybean endogenous genes and the five transgenic soybean lines had good specificity. There was no cross amplification or non-specific amplification between the primers, and the detection sensitivity of the multiplex PCR method reached 0.1%. The method can be used as an aid to the detection of genetically modified soybeans and their product components, and can quickly detect the corresponding strains in the transgenic soybean and its products.