抗草甘膦转基因玉米外源基因ddPCR拷贝数分析

发布时间：2019-05-07 03:50

【摘要】：【目的】本研究是从前期获得的100余份转基因抗除草剂草甘膦玉米品系中,挑选T抗-1等14个转基因玉米品系进行拷贝数检测。【方法】采用一种新型的拷贝数分析方法——微滴数字PCR技术(droplet digital PCR,简称dd PCR),设计特异引物对外源抗草甘膦基因2m G2-epsps进行绝对定量分析。【结果】供试的14份转基因材料中10份是单拷贝品系。同时,通过引物探针特异性试验、叶片基因组DNA的PCR抑制和浓度检测、转基因玉米的外源基因2m G2-epsps的实时PCR检测和基因组DNA的酶切等系列研究,建立稳定的转基因玉米拷贝数分析的dd PCR检测体系。【结论】微滴数字PCR技术为这批转基因玉米的下一步转基因生物安全评价提供了重要参数,同时建立了转基因玉米(genetically modified maize)外源基因拷贝数dd PCR分析方法。
[Abstract]:[objective] in this study, more than 100 transgenic herbicide-resistant glyphosate-resistant maize lines were obtained. Fourteen transgenic maize lines, such as T-resistance-1, were selected for copy number detection. [methods] A new copy number analysis method, microdrop digital PCR technique (droplet digital PCR,), was used to detect copy number of transgenic maize lines (dd PCR), for short). Specific primers were designed for absolute quantitative analysis of exogenous glyphosate resistance gene 2m G2-epsps. [results] of the 14 transgenic materials tested, 10 were single copy lines. At the same time, PCR inhibition and concentration detection of genomic DNA in leaves, real-time PCR detection of exogenous gene 2m G2-epsps and restriction enzyme digestion of genomic DNA in transgenic maize were carried out by primer probe specificity test. A stable dd PCR detection system for copy number analysis of transgenic maize was established. [conclusion] the microdrop digital PCR technique provides important parameters for the next step of transgenic biological safety assessment of these transgenic maize. At the same time, the dd PCR analysis method of foreign gene copy number of transgenic maize (genetically modified maize) was established.
【作者单位】：四川省农业科学院生物技术核技术研究所;中国农业科学院生物技术研究所;
【基金】：四川省农科院高新技术研究应用专项(2016GXTZ-001);四川省农科院财政基因工程项目(2016ZYPZ-003) 转基因生物新品种培育重大专项(2014ZX0800301B)
【分类号】：S513

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