定性PCR方法检测转基因玉米MON810转化事件的研究  

Event-specific Detection Method of Transgenic Maize MON810 by Using Qualitative Polymerase Chain Reaction(PCR)

作　　者：易小平[1,2] 贺萍萍[1,2] 夏启玉[1,2] 肖苏生[1,2] 谢翔[1,2] 杨小亮[1,2] 李美英[1,2] 郭安平[1,2] 

机构地区：[1]中国热带农业科学院热带生物技术研究所农业部热带作物生物技术与遗传资源利用重点实验室,海南海口571101 [2]农业部转基因植物及植物用微生物环境安全监督检验测试中心,海南海口571101

出　　处：《热带作物学报》2014年第12期2384-2390,共7页Chinese Journal of Tropical Crops

基　　金：南繁区生物安全监测预警及控制关键技术研究与示范(No.201403075);海南南繁区Bt水稻基因飘移风险监测与控制(No.2014ZX08012004-011);海南省自然科学基金(No.311082)

摘　　要：转基因玉米MON810(Yield Gard R)是孟山都公司通过DNA重组技术和微注射轰击研发的一种具有对欧洲玉米螟(ECB;Ostrinia nublialis)有特殊抗性的转基因玉米品系,目前在世界各地已经得到广泛种植,为加强对该品系玉米的安全管理,本研究旨在建立MON810品系玉米的转化事件特异性定性PCR检测方法。根据MON810的插入序列信息,在3′端的侧翼序列处设计定性PCR检测的引物,检测MON810在其他几种常见转基因作物混合样品的特异性。结果表明,该方法具有很好的特异性。同时检测该引物系统的扩增灵敏度,结果表明,检测引物的灵敏度可达0.1%。建立的MON810特异定性PCR检测方法经全国7家实验室的验证,进一步证实该方法能够特异地检测出样品中的MON810转化事件,检测方法的灵敏度可达0.1%,且检测结果具有良好的可重复性和可重现性。MON810转化事件定性PCR检测方法的建立可满足于抗虫转基因玉米MON810及其衍生品种生物安全管理的需要。Genetically modified maize（Zea mays）MON810（YieldGard（R））was developed by Monsanto Company through recombinant DNA and micro injection bombardment technology,which is resistant to European corn borer （ECB; Ostrinia nublidis）specially.It is widely planted in the world.The aim of the study is to develop an event-specific qualitative detection method to manage the spreading of MON810 in China.Based on the information of the inserted sequences of MON810,a pair of primers werer designed in the 3＇end of the flanking binding site.No amplification product was detected in several transgenic crop common mixed samples and the limit detection was the mass fraction of 0.1％ MON810 maize,indicating the method could be used specifically and sensitively.The established method was then validated by another 7 external laboratories,the results showed that the method could be specifically detect the samples of MON810 with the detection sensitivity about the mass fraction of 0.1％ MON810 maize,and also indicating the method had a good repeatability and reproducibility.So the method of detection MON810 can be suitable to the biosafety management of MON810 and its derivates.

关 键 词：转基因玉米MON810 定性PCR 灵敏度 特异性 

分 类 号：S19[农业科学—农业基础科学]