转基因玉米CM8101实时荧光定量PCR检测方法的建立  

Establishment of qRT-PCR Method of Detection Transgenic Maize(Zea mays)CM8101

作　　者：龙丽坤[1] 赵宁[1] 李葱葱[1] 何禹璇 董立明[1] 闫伟[1] 李飞武[1] LONG Li-Kun;ZHAO Ning;LI Cong-Cong;HE Yu-Xuan;DONG Li-Ming;YAN Wei;LI Fei-Wu(Institute of Agricultural Quality Standards 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：《农业生物技术学报》2021年第5期1007-1015,共9页Journal of Agricultural Biotechnology

基　　金：国家转基因生物新品种培育重大专项(2018ZX08012001-003)。

摘　　要：CM8101转基因玉米(Zea mays)是我国自主研发的具有产业化前景的转基因玉米品系,对CM8101转基因玉米及其产品进行定量检测方法的研发,是转基因作物安全监管的技术要求。本研究根据转基因玉米CM81015’端旁侧序列信息,设计实时荧光定量引物和Taqman探针,经特异性、线性、检测限、定量限、准确性、精确性和重复性等测试,建立了CM8101玉米转化体特异性实时荧光PCR定量检测方法。样品测定结果表明,本定量检测方法特异性强、准确性好,可以对低至0.1%的CM8101转基因成分进行准确定量,检测限可达到10个拷贝。本研究基于qRT-PCR技术,开发了CM8101转基因玉米的准确定量检测方法,该方法有望为CM8101转基因玉米的安全监管提供必要的技术支持。Genetically modified maize(Zea mays)CM8101 is one of the transgenic maize lines with commercial prospect developed independently in China.The research of quantitative detection method for CM8101 transgenic maize and its products is one of the technical requirements for the safety supervision of transgenic crops.In this study,real-time fluorescence quantitative primers and TaqMan probe were designed according to the 5’side sequence information of transgenic maize CM8101.After specificity,linearity,limit of detection,limit of quantification,accuracy,precision and reproducibility tests,a specific real-time fluorescence PCR quantitative detection method for CM8101 maize was established.The results of sample determination showed that this quantitative method had strong specificity and good accuracy,and could accurately quantify CM8101 transgenic components as low as 0.1%,and the detection limit could reach to 10 copies.In this study,an accurate quantitative detection method for CM8101 transgenic maize was developed based on qRT-PCR technology,which is expected to provide necessary technical support for CM8101 transgenic corn safety supervision.

关 键 词：转基因玉米 CM8101 特异性 实时荧光PCR 定量检测 

分 类 号：S188[农业科学—农业基础科学]