转基因抗虫耐除草剂玉米C0030.1.1定性PCR检测方法的建立  

Development of Detection Method of Insect-Resistant and Herbicide-Tolerant Maize C0030.1.1

作　　者：董文凤 梁晋刚 李夏莹 刘鹏程 张旭冬 王颢潜 陈子言 张秀杰 DONG Wenfeng;LIANG Jingang;LI Xiaying;LIU Pengcheng;ZHANG Xudong;WANG Haoqian;CHEN Ziyan;ZHANG Xiujie(Development Center of Science and Technology,Ministry of Agriculture and Rural Affairs,Beijing 100176,China)

机构地区：[1]农业农村部科技发展中心,北京100176

出　　处：《东北农业科学》2023年第2期49-54,63,共7页Journal of Northeast Agricultural Sciences

基　　金：农业农村部转基因生物新品种培育科技重大专项(2016ZX08012003,2019ZX08012-004)。

摘　　要：转基因抗虫耐除草剂玉米C0030.1.1是由北京大北农生物技术有限公司自主研发的转cry1Ab基因和epsps基因抗虫耐除草剂玉米转化体。为了建立转基因抗虫耐除草剂玉米C0030.1.1的检测方法,本研究在外源基因插入位点的左、右边界上设计引物,通过引物筛选、特异性测试、引物浓度和退火温度测试、灵敏度测试等试验,最终在右边界建立了抗虫耐除草剂玉米C0030.1.1的定性PCR检测方法,灵敏度为0.1%。8家单位循环验证结果表明本方法具有很好的重复性和再现性,可以特异性检测到转化事件。本研究建立的方法适用于产品中含有C0030.1.1玉米转基因成分的检测。Genetically modified(GM)maize(Zea mays L.)C0030.1.1 is developed by Beijing DaBeiNong Biotechnology Co.,Ltd.The purpose of this study is to develop the event-specific qualitative detection method of this maize C0030.1.1.Based on the foreign inserted gene,a series of specific PCR primers were designed on the left and right of the flanking binding site.And then the PCR reaction systems were screened and optimized based on primer screening,specificity test,sensitivity test,differential primer concentration and a series of annealing temperatures.The maize C0030.1.1 events-specific qualitative PCR detection method was established finally on the right of the flanking binding site.The detection sensitivity was 0.1%.After validated by 8 external laboratories,results showed that this method could specifically detect the samples of C0030.1.1 event and had a good repeatability and reproducibility.

关 键 词：转基因玉米 定性PCR C0030.1.1 

分 类 号：S513[农业科学—作物学]