抗虫耐除草剂玉米GH5112E-117C定性PCR检测方法  

Qualitative PCR Assay for the Detection of GH5112E-117C Transgenic Maize Resistant to Insects and Herbicides

作　　者：李葱葱[1] 谢苹[1] 董立明[1] 夏蔚[1] 兰青阔 闫伟[1] 龙丽坤[1] 李飞武[1] LI Cong-cong;Xie Ping;DONG Li-ming;XIA Wei;LAN Qing-kuo;YAN Wei;LONG Li-kun;LI Fei-wu(Institute of Agricultural Quality Standard and Testing Technology,Jilin Academy of Agricultural Sciences,Changchun 130033;Tianjin Institute of Agricultural Quality Standard and Testing Technology,Tianjin 300381)

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 [2]天津市农业质量标准与检测技术研究所,天津300381

出　　处：《生物技术通报》2020年第5期64-67,共4页Biotechnology Bulletin

基　　金：转基因重大专项课题(2018ZX08012-001)。

摘　　要：转基因抗虫耐除草剂玉米GH5112E-117C是由北京奥瑞金公司研发的转mG2-aroA基因和mcry1Ah基因抗虫耐除草剂玉米新品系,对玉米螟、粘虫等鳞翅目害虫具有抗性、耐草甘膦除草剂的转基因玉米,在我国具有重要产业化应用前景。本文研发了GH5112E-117C转基因玉米的定性PCR检测方法,该方法能特异性地检测转基因抗虫耐除草剂玉米GH5112E-117C,在每个反应引物浓度为0.2 μmol/L、Taq DNA 聚合酶0.625 U、DNA模板50 ng,退火温度为58℃的条件下,检测灵敏度可稳定达到0.1%。该方法为转基因抗虫耐除草剂玉米GH5112E-117C的精准检测提供了一种新的技术手段,为农业转基因监管提供技术支撑。Transgenic maize GH5112E-117C is a new strain with mG2-aroA gene and mcry1Ah genes and developed by Beijing Origen Company.It is resistant to lepidopteran pests such as corn borer and armyworm,and it is tolerant to glyphosate-herbicide.Thus this strain has important industrial application prospects in China.A qualitative PCR assay for transgenic maize GH5112E-117C was developed in this paper.The assay had promising specificity to detect the transgenic maize GH5112E-117C with resistance to insects and tolerance to herbicide.At a concentration of 0.2 μmol/L primer,TaqDNA polymerase 0.625 U,DNA template 50 ng,and an annealing temperature of 58℃,the detection sensitivity of the assay stably achieved to 0.1%.This assay provides a new technical means for accurately testing insect-and herbicide-resistant transgenic maize GH5112E-117C,which provides the technical support for transgenic supervision in agriculture.

关 键 词：转基因玉米 转化事件 PCR 检测方法 

分 类 号：S435.132[农业科学—农业昆虫与害虫防治]