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3mmol/L、反应时间为45min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 and3 mmol/L Mg2+.The method developed in this paper could specifically detect glyphosate-resistant soybeans.And the low d...
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