摘要
以转基因水稻中最常用的CaMV35S启动子、NOS终止子、Cry1Ab/Ac基因、HPT基因及SPS水稻内标基因为研究对象,利用5种不同的荧光信号(FAM、HEX、Taxas Red、Cy5、Cy5.5)进行多重实时聚合酶链式反应(realtime polymerase chain reaction,real-time PCR)检测方法的研究。通过引物组合筛选、反应体系优化、特异性测试、灵敏度测试、适用性测试等一系列实验,建立了5重real-time PCR方法,灵敏度可达0.032%。此方法具有灵敏度高、结果准确、通量大等优点,可实现水稻中转基因成分的快速、高效检测。
In this study,based on the CaMV35S promoter,the NOS terminator,Cry1Ab/Ac,HPT and the rice endogenous gene SPS,five different fluorescent probes(FAM,HEX,Taxas Red,Cy5,and Cy5.5)were used to develop and optimize a pentaplex real-time polymerase chain reaction(real-time PCR)method to detect genetically modified rice.For this purpose,a series of experiments was carried out to screen primer combinations,optimize the reaction system,and evaluate the specificity,sensitivity and applicability.The sensitivity of the proposed method was 0.032%.This method has the characteristics of high sensitivity,accuracy and high throughput and allows the rapid and efficient detection of genetically modified components in rice.
作者
董立明
杨帆
邢珍娟
李葱葱
闫伟
龙丽坤
李飞武
DONG Liming;YANG Fan;XING Zhenjuan;LI Congcong;YAN Wei;LONG Likun;LI Feiwu(Institute of Agricultural Quality Standard and Testing Technology,Jilin Academy of Agricultural Sciences,Changchun 130033,China)
出处
《食品科学》
EI
CAS
CSCD
北大核心
2021年第24期329-334,共6页
Food Science
基金
吉林省农业科技创新工程项目(CXGC2017TD011)
国家转基因生物新品种培育重大专项(2018ZX08012001)。
关键词
转基因水稻
筛查元件
单一实时聚合酶链式反应
5重实时聚合酶链式反应
高通量检测方法
genetically modified rice
screening element
single real-time polymerase chain reaction
pentaplex real-time polymerase chain reaction
high-throughput detection method
