转基因大豆BPS—CV127-9PCR定量检测研究  

Quantitative Detection of Genetically Modified Soybean BPS-CV127-9

作　　者：李宁[1,2] 赵蕾[3] 孙红炜[1] 李凡[1] 杨淑珂[1] 路兴波[1] 

机构地区：[1]山东省农业科学院植物保护研究所/山东省植物病毒学重点实验室,山东济南250100 [2]济宁医学院公共卫生学院,山东济宁272067 [3]山东师范大学生命科学学院,山东济南250014

出　　处：《大豆科学》2012年第5期808-812,共5页Soybean Science

摘　　要：采用SYBRGreen实时荧光定量PCR技术，建立转基因大豆BPS—CV127-9的定量检测方法。通过设计特异引物，扩增内标准基因1ectin和BPS．CV127-9的5’侧翼序列，建立2种基因的拷贝数-CT标准曲线，根据标准曲线方程计算样品中的转基因含量，并且通过熔解曲线分析扩增反应特异性。结果表明，1ectin基因和侧翼序列标准曲线线性关系良好，酽值分别为0．999和0．998，变异系数（CV）1．50％～18．51％、标准偏差（SD）0．02～0．07。检测4个已知BPS．CV127-9含量（1％、0．5％、0．1％、0．05％）的转基因混合样品，实测值与实际值接近。该检测方法具有快速、灵敏、准确、特异、高通量等优点，可以作为转基因大豆BPS—CV127-9的定量检测方法。A quantitative method to detect the transgenic soybean BPS-CV-127-9 by using real-time PCR technique based on fluorescence dye SYBR Green I was investigated in this study. The endogenous lectin gene and the 5＇ flanking sequences of BPS-CV-127-9 were amplified through the specific primers, and the transgenic content was then calculated according to the standard curve equation. Meanwhile the specificity of PCR amplification was analyzed by corresponding melting curves. The re- suits showed that the standard curves of lectin and the 5＇ flanking sequences of BPS-CV-127-9 genes have good linear relation- ship, and their R2 values were 0.999 and 0.998, respectively. The coefficient of variance was 1.50% -18.51% and standard deviation was 0.02-0.07. Four mixed samples with genetically modified contents of BPS-CV-127-9 was 0.05% ,0.1% ,0.5% and 1% respectively were detected, and the detection results agreed well with actual value. In conclusion, this method was fast, sensitive,simple, accurate, specific and of high throughput, and could be used to detect transgenic soybean BPS-CV-127-9 quantificationally.

关 键 词：品系特异性 实时荧光定量PCR 转基因大豆 

分 类 号：S816.35[农业科学—饲料科学]