原创《转基因耐除草剂玉米VCO—O—5定性PCR标准检测方法建立》：这是一篇与除草剂论文范文相关的免费优秀学术论文范文资料,为你的论文写作提供参考。
摘 要 转基因耐除草剂玉米VCO-O/1981-5是由加拿大Genective SA公司通过农杆菌侵染幼胚转化获得的1种新的抗草甘膦除草剂转基因玉米转化体,目前已经进行商业化生产应用.本研究的目的是建立转基因VCO-O/1981-5玉米品系特异性转化事件标准化定性PCR（polymerase chain reaction,PCR）检测方法.根据VCO-O/1981-5玉米插入的外源基因和左、右侧翼序列设计了一系列特异性的PCR引物,分别进行了引物筛选和PCR反应体系优化,建立了耐除草剂玉米VCO-O/1981-5转化体特异性定性PCR标准化检测方法,对建立的检测方法在非转基因VCO-O/1981-5玉米的混合样品中进行了特异性测试,仅从含有VCO-O/1981-5玉米样品中扩增到目的条带,表明检测方法具有很好的特异性,检测的灵敏度为0.1%.建立的检测方法经过实验室内和实验间循环验证比对,具有良好的重复性和再现性.本研究建立的VCO-O/1981-5玉米定性PCR标准化方法适用于产品中含有VCO-O/1981-5玉米转基因成分的检测.
关键词 VCO-O/1981-5；定性PCR；灵敏度；品系特异性；标准化检测方法
中图分类号 S19 文献标识码 A
Abstract Transgenic maize（Zea mays）VCO-O/1981-5, developed by Canada Genective SA Company through agrobacterium mediated transformation of immature embryos technology is commercially produced. The aim of this study is to develop an event-specific and standardized qualitative polymerase chain reaction（PCR）detection method for herbicide-tolerant maize VCO-O/1981-5. Based on the foreign gene inserted, and the right and left flanking sequences of the gene, a series of specific PCR primers were designed on the right and left of the flanking binding site. And then the PCR reaction systems were screened and optimized based on differential primer concentration and a series of annealing temperatures from 50 ℃ to 60 ℃. Finally, the developed event-specific and standardized PCR detection method was tested in several mixed non-transgenic samples. The results showed that no amplification product was found in the non-VCO-O/1981-5 transgenic mixture. The limit detection sensitivity was the mass fraction of 0.1% VCO-O/1981-5 maize. The developed method was then validated by another 7 external laboratories, and the results showed that this method could be specifical to detect the samples of VCO-O/1981-5 with the detection sensitivity about the mass fraction of 0.1% VCO-O/1981-5 maize, and also indicated the method had a good repeatability and reproducibility. So the method of detection VCO-O/1981-5 should be suitable to the biosafety management of VCO-O/1981-5 and its derivate.
Key words VCO-O/1981-5； qualitative polymerase chain reaction（PCR）； detection sensitivity； specifically
doi 10.3969/j.issn.1000-2561.2017.12.019
转基因耐除草剂玉米VCO-O/ 1981-5是由加拿大Genective SA公司通过农杆菌侵染幼胚转化获得的一种新的抗草甘膦除草剂转基因玉米转化体.VCO-O/ 1981-5玉米含有epsps grg23ace5外源基因,编码修饰的5-烯醇丙酮莽草酸-3-磷酸合酶（5-enolpyruvylshikimate-3-phosphate synthase,EPSPS）,该基因从球形节杆菌分离,EPSPS对草甘膦具有天然的抗性[1].VCO-O/ 1981-5玉米自2013年起已經获得在美国和加拿大等国家的商业化应用批准许可,并进行了大面积商业化种植生产应用[1], 迄今为止,我国尚未发布专门针对耐除草剂玉米VCO-O/ 1981-5及其衍生品种检测的标准化定性PCR检测方法,因此对转基因耐除草剂玉米VCO-O/ 1981-5定性PCR检测方法标准化研究,将为VCO-O/ 1981-5转基因玉米批准进口到中国后的安全监管提供理论依据,具有重要的现实意义.
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结论：转基因耐除草剂玉米VCO—O—5定性PCR标准检测方法建立为关于本文可作为相关专业除草剂论文写作研究的大学硕士与本科毕业论文自制除草剂2天见效论文开题报告范文和职称论文参考文献资料。
