定性PCR技术及卡那霉素对8个番茄品种的转基因检测

GMO Detection in Eight Different Tomato Varieties by Using Qualitative PCR Technique and Kanamycin-Resistance

本研究利用定性PCR技术对8个不同番茄品种中外源基因35S启动子、NPTⅡ基因和NOS终止子进行检测。实验结果表明,阳性对照可以稳定扩增到预期的大小片段,而待检测品种和阴性对照则没有扩增到预期产物。利用100mg/L的卡那霉素对8个不同番茄品种的种子进行萌发试验,再用50mg/L的卡那霉素对番茄外植体(子叶和下胚轴)进行抗性验证,结果显示卡那霉素抗性筛选实验结果与定性PCR结果一致。本文通过对这两种检测方法的优缺点进行研究分析及比较,初步建立了一个对不同品种番茄进行转基因检测的体系。

In this article, the exogenous gene 35S promoter, NPT Ⅱ gene and NOS terminator in eight different tomato varieties can be detected by PCR amplification method. The results showed that positive control can be amplified stable the expected fragment while under-test varieties and negative control can not be amplified. At the same time, the Kanamycin resistance of these tomato varieties were sprouting test, the seeds of these varieties were inoculated to test their germination under 100 mg/L Kanamycin, then the resistance of their explants can be further defined under 50 mg/L Kanamyein. The result of Kanamycin resistance was consistent with that of PCR. In this paper, transgenic detection system of different tomato varieties has been established preliminarily by using compared to advantages and disadvantages between two detection methods.