| 朱琳峰, 彭焕, 黄文坤, 张东升, 孔令安, 彭德良*.抗草甘膦转基因大豆Cp4-epsps基因快速简便的LAMP 检测方法[J].植物保护,2015,41(3):86-92. |
| 抗草甘膦转基因大豆Cp4-epsps基因快速简便的LAMP 检测方法 |
| Rapid and simple detection of Cp4-epsps gene in glyphosate-resistant soybean by loop-mediated isothermal amplification |
| 投稿时间：2014-04-11 修订日期：2014-06-25 |
| DOI： |
| 中文关键词: 抗草甘膦转基因大豆 Cp4-epsps LAMP 快速检测 |
| 英文关键词:glyphosate-resistant soybean Cp4-epsps LAMP rapid and simple detection |
| 基金项目:转基因生物新品种培育科技重大专项(2014ZX008011-003) |
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| 中文摘要: |
| 针对抗草甘膦转基因大豆的外源基因Cp4-epsps, 建立了一种基于环介导等温扩增（loop-mediated isothermal amplification, LAMP）技术的抗草甘膦转基因大豆的检测体系, 其扩增产物既可利用常规琼脂糖凝胶电泳检测, 还可通过SYBR Green I染色进行快速检测。LAMP 检测体系中dNTPs浓度为0.8 mmol/L、Mg2+浓度为3 mmol/L、反应时间为45 min时扩增效果最佳, 其检测灵敏度为5μg/L, 比常规PCR灵敏100倍。田间实际检测结果表明, LAMP 检测结果和PCR检测结果完全一致, 准确率为100%。本研究所建立的抗草甘膦转基因大豆LAMP 检测方法具有简便快速、特异性强、灵敏度高等特征, 是一种能够用于抗草甘膦转基因大豆检测、田间基因漂移监测和环境安全研究的有力工具。 |
| 英文摘要: |
| The LAMP (loop-mediated isothermal amplification) assay based glyphosate-resistant soybean detection system was developed in this paper. A set of four basic primers and two loop primers was designed to target the exogenous Cp4-epsps gene. The LAMP products were detected by agarose gel electrophoresis and SYBR Green I. The optimum result was obtained at 45 min of reaction time, 0.8 mmol/L dNTPs and 3 mmol/L Mg2+. The method developed in this paper could specifically detect glyphosate-resistant soybeans. And the low detect limit of genomic DNA of glyphosate-resistant soybeans was 5 μg/L which was 100 times higher than that of regular PCR. The field testing showed that the results of LAMP detection were consistent with those of PCR, and the accuracy of this method was 100%. The LAMP method we developed for glyphosate-resistant soybean detection is rapid, efficient, specific, sensitive and convenient, which make it a useful tool for detection of glyphosate-resistant soybean, monitoring of gene flow and research on environmental security. |
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