高灵敏度电化学发光PCR方法检测花椰菜花叶病毒35 S启动子

大多数转基因植物中使用花椰菜花叶病毒(cauliflower mosaic virus,CaMV)35 S作为启动子,因此可通过检测该启动子来判断植物样品中是否含有转基因成分。实验将高灵敏度电化学发光PCR方法用于检测转基因烟草中的CaMV35 S启动子,将该启动子的PCR产物与生物素标记的探针杂交,可以起到特异性筛选产物的作用;与发光标记物——三联吡啶钌标记的探针杂交,从而实现电化学发光检测。两种探针同时与待测样品的PCR产物进行杂交,进一步对样品进行特异性筛选,从而提高了检测的准确性,避免了假阳性结果的产生。实验结果表明:该法可以准确的区分待测样品中是否含有35 S启动子,从而区别转基因烟草和非转基因烟草。电化学发光PCR方法灵敏度高,可靠性强,操作简便,结果准确,有望成为一种高效的转基因植物检测方法。

Highly Sensitive Electrochemiluminescence-PCR Method for the Detection of CaMV 35 S Promoter

Most genetically modified(GM) plants contain cauliflower mosaic virus 35 S(CaMV 35 S) promoter.So,whether the plants contain GM component can be discriminated by detecting the CaMV 35 S promoter.In this study,the highly sensitive electrochemiluminescence-polymerase chain reaction(ECL-PCR) assay combined with two types of nucleic acid probes hybridization was applied to detect CaMV 35 S promoter in tobaccos.The experiment results show that the CaMV 35 S promoter can only be detected in GM tobaccos,thus,the GM tobaccos and the non-GM tobaccos can be clearly discriminated.The method may provide a new means for the detection of GM plants due to its sensitivity,simplicity and high efficiency.