[关键词]
[摘要]
通过对我国批准进口的和获得农业转基因生物安全证书的转基因玉米转化体的序列分析发现,除DAS40278-9和BLVA430101外,CaMV35s启动子、NOS终止子、Cry1Ab/Ac基因和pat基因覆盖了21种玉米转基因转化体。通过引物组合筛选、反应体系优化、灵敏度测试、适用性测试等实验,建立了基于4个通用筛选元件及zSSIIb内源基因的五重荧光PCR和基于转化体特异性序列的二重荧光PCR检测转基因成分方法体系。方法参数测定结果表明,此方法特异性强、稳定性好,检测灵敏度达到0.05%。同时,此方法体系不仅适用于转基因玉米成分筛查,对大豆、水稻等多种作物进行转基因成分筛选鉴定也有良好的适用性。
[Key word]
[Abstract]
Through sequence analysis of genetically modified corn transformants approved for import and obtained agricultural genetically modified organisms safety certificates in China, it was found that CaMV35s promoter, NOS terminator, Cry1Ab/Ac gene, and Pat gene covered 21 corn transgenic events. In this study, a series of experiments, including primer combination screening, reaction system optimization, sensitivity test, and applicability test were conducted. Consequently, a five-plex fluorescent PCR system and a duplex fluorescent PCR system were established based on four common screening elements and an endogenous gene zSSIIb. The measurement results of parameters showed that these two multiplex fluorescent PCR detection systems established in this study are characterized by strong specificity, good stability high sensitivity, which can reach 0.05%. Meanwhile, these two systems can also be used to screen and identify genetically modified ingredients in soybean, rice and many other crops.
[中图分类号]
[基金项目]
国家转基因生物新品种培育科技重大专项(2018ZX08012-001)