摘要
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。
The soybean lines 305423, MON89788, CV127 and GTS40-3-2 are the most widely commercialized genetically modified soybeans and account for more than 80% of the world total transgenic soybean planting acreage. In this study, the specific primers were designed according to the border sequences of five GM soybean events. By verifying the applicability, specificity and sensitivity of the primers the multiplex PCR conditions,including the amount of primers and the annealing temperature, were optimized. Then a multiplex PCR detection system which can be used to amplify soybean lectin gene and introduced genes in five GM soybean events simultaneously was developed. The results showed that the primers applied in this method had good specificity, no cross-amplification and nonspecific amplification. The detection limit of this method could reach 0. 1%. This method could be used as an auxiliary means to detect the genetically modified ingredients rapidly.
作者
董立明
李葱葱
邢珍娟
邵改革
夏蔚
闫伟
李飞武
DONG Li-ming LI Cong-cong XING Zhen-juan SHAO Gai-ge XIA Wei YAN Wei LI Fei-wu(Institute of Agricultural Quality Standard and Testing Technology, Jilin Academy of Agricultural Sciences, Changchun ! 30033, Chin)
出处
《大豆科学》
CAS
CSCD
北大核心
2016年第6期1002-1007,共6页
Soybean Science
基金
国家转基因生物新品种培育重大专项(2014ZX0801202B)
吉林省农业科技创新工程项目(2013)
关键词
转基因大豆
多重PCR
品系特异性检测
Genetically modified soybean
Multiplex PCR
Event-specific detection
