| 转基因耐除草剂玉米C0010.2.2定性PCR检测方法的建立 |
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| 引用本文： | 董文凤,梁晋刚,李夏莹,刘鹏程,张旭冬,王颢潜,陈子言,张秀杰.转基因耐除草剂玉米C0010.2.2定性PCR检测方法的建立[J].玉米科学,2021,29(5):57-64. |
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| 作者姓名： | 董文凤 梁晋刚 李夏莹 刘鹏程 张旭冬 王颢潜 陈子言 张秀杰 |
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| 作者单位： | 农业农村部科技发展中心, 北京 100176 |
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| 基金项目： | 转基因生物新品种培育科技重大专项（2016ZX08012003，2019ZX08012-004） |
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| 摘 要： | 转基因耐除草剂玉米C0010.2.2是北京大北农生物技术有限公司利用农杆菌介导法,将epsps基因和pat基因转到受体玉米DBN567获得的耐除草剂玉米转化体,具有耐除草剂草甘膦和草铵膦性状。研究建立转基因耐除草剂玉米C0010.2.2的检测方法,在外源基因插入位点的左、右边界分别设计引物,经过引物筛选、特异性测试、灵敏度测试、退火温度和引物浓度测试,建立转基因耐除草剂玉米C0010.2.2的定性PCR检测方法,该方法的检出限和灵敏度可达到0.1%。验证结果表明,该方法可以特异性检测到转化事件,具有很好的重复性和再现性。
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| 关 键 词： | 玉米 C0010.2.2 定性PCR |
| 收稿时间： | 2020/9/8 0:00:00 |
Development of Detection Method of Genetically Modified Herbicide-tolerant Maize C0010.2.2 |
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| DONG Wen-feng,LIANG Jin-gang,LI Xia-ying,LIU Peng-cheng,ZHANG Xu-dong,WANG Hao-qian,CHEN Zi-yan,ZHANG Xiu-jie.Development of Detection Method of Genetically Modified Herbicide-tolerant Maize C0010.2.2[J].Journal of Maize Sciences,2021,29(5):57-64. |
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| Authors: | DONG Wen-feng LIANG Jin-gang LI Xia-ying LIU Peng-cheng ZHANG Xu-dong WANG Hao-qian CHEN Zi-yan ZHANG Xiu-jie |
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| Institution: | Development Center of Science and Technology, Ministry of Agriculture and Rural Affairs, Beijing 100176, China |
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| Abstract: | Genetically modified maize(Zea mays L.) C0010.2.2 was developed by Beijing DaBeiNong Biotechnology Co., Ltd. through Agrobacterium-mediated transformation which transformed epsps and pat gene to maize DBN567 characterized by resistance to glyphosate and glufosinate. The purpose of this study is to develop the qualitative detection method of genetically modified maize C0010.2.2. A series of specific PCR primers were designed on the right and left of the foreign gene inserted binding site, and a qualitative detection method for genetically modified herbicide-tolerent maize C0010.2.2 was established according to primer screening, specificity testing, sensitivity testing, primer concentration testing and annealing temperatures testing. The method was verified that has a good specificity and repeatability with the limit of detection(LOD) and sensitivity 0.1% by 8 external laboratories. |
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| Keywords: | Maize C0010 2 2 Qualitative PCR |
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