| 摘要: |
| 试验建立大豆中转基因成分的定量测定方法。采用双链DNA SYBR Green I结合染料实时荧光定量PCR技术，扩增编码大豆凝集素的内参照基因lec和抗草甘膦转基因大豆中的外源花椰菜花叶病毒CaMV35s基因。绘制2种基因扩增的循环数一拷贝数标准曲线图，根据标准曲线方程计算样品中的转基因含量；并作重现性和熔解曲线分析。结果表明，lec和CaMV35s基因标准曲线方程线性好，R^2值分别达到0．9997和0．9992。不同转基因含量的标准及模拟大豆样品定量显示，实测值与实际值接近，相对偏差5％～11％。SYBR Green I实时定量PCR方法可用于定量测定大豆中转基因品种的含量。 |
| 关键词: 实时定量PCR SYBR Green I荧光染料 抗草甘膦大豆 转基因标识 |
| DOI：10.11841/j.issn.1007-4333.2005.03.063 |
| 修订日期：2004-11-15 |
| 基金项目:农业部专项基金资助项目(农技函[2001]184号)，饲料工业应用HACCP体系的关键技术研究(6031002) |
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| Quantitative PCR method for detecting glyphosate- tolerant gene transfer in soybeans |
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| Abstract: |
| Quantitative determination of the transgenic component in soybeans was developed in this study. Based on real-time quantitative PCR technique with SYBR Green I as fluorescent dye combined to double-strand DNA, the endogenous lec gene (encoding soybean lectin) and exotic CaMV35s gene (encoding 35S promotor of CaMV inserted in glyphosate-tolerant soybeans) were amplified. The transgenic ratio was then calculated according to the standard Ct-copies linear graphs of these two genes. The reproducibility and melting curves of the genes were also analyzed. The results showed that the standard equations of lec and CaMV35s genes had higher R2 value of 09997 and 09992 respectively. The difference between the determined and the known transgenic levels of standard and simulated soybeans was only 5%-11%. It was thus suggested that the PCR method with fluorescent dye SYBR Green I was suitable for quantifying the weight ratio of transgenic soybeans in soybean products. |
| Key words: real-time quantitative PCR,fluorescence dye SYBR Green I,glyphosate-tolerant soybeans,transgenic labeling |
