应用多重荧光PCR快速筛查作物中转基因成分研究  

Rapid Screening of Genetically Modified Ingredient in Crops by Multiplex Real-time PCR

作　　者：邢珍娟[1] 董立明[1] 龙丽坤[1] 刘娜[1] 夏蔚[1] 谢彦博 李葱葱[1] 李飞武[1] XING Zhen-juan;DONG Li-ming;LONG Li-kun;LIU Na;XIA Wei;XIE Yan-bo;LI Cong-cong;LI Fei-wu(Institute of Agricultural Quality Standard and Testing Technology,Jilin Academy of Agricultural Sciences,Changchun 130033,China)

机构地区：[1]吉林省农业科学院农业质量标准与检测技术研究所,长春130033

出　　处：《玉米科学》2021年第4期35-42,共8页Journal of Maize Sciences

基　　金：国家转基因生物新品种培育科技重大专项(2018ZX08012-001)。

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。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.

关 键 词：玉米 转基因作物 多重实时荧光PCR 快速筛查 

分 类 号：S513.037[农业科学—作物学]