LI Ning,ZHAO Lei,SUN Hong-wei,et al.Quantitative Detection of Genetically Modified Soybean BPS-CV127-9[J].Soybean Science,2012,31(05):808-812.[doi:10.3969/j.issn.1000-9841.2012.05.026]
转基因大豆BPS-CV127-9 PCR定量检测研究
- Title:
- Quantitative Detection of Genetically Modified Soybean BPS-CV127-9
- 文章编号:
- 1000-9841（2012）05-0808-05
- Keywords:
- Event-special; Real-time PCR; Genetically modified soybean
- 分类号:
- S565.1
- 文献标志码:
- A
- 摘要:
- 采用SYBR Green 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lectin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，lectin基因和侧翼序列标准曲线线性关系良好，R2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。
- Abstract:
- 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 results showed that the standard curves of lectin and the 5′ flanking sequences of BPS-CV-127-9 genes have good linear relationship,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.
参考文献/References:
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备注/Memo
基金项目：国家转基因重大专项(2008ZX08012-001)。