论文部分内容阅读
本研究利用病毒诱导基因沉默技术对转Bt基因棉花选择标记基因nptⅡ进行沉默,分析标记基因转录后沉默的可行性。以烟草脆裂病毒载体为骨架构建nptⅡRNAi载体,利用农杆菌浸染棉花子叶,获得nptⅡ干涉的转Bt基因棉花植株,然后涂抹卡那霉素进行检测。结果表明nptⅡ沉默植株叶片出现黄斑症状,RT-PCR和q RT-PCR检测表明叶片、根和茎中nptⅡ基因转录分别被抑制了99.25%、99.05%和98.65%,沉默后期的转录抑制也达到98%。本研究结果为解决已经在环境中释放转基因生物发生意外扩散和不可预料的生物安全事件提供了快速有效的应对方法和新思路。
In this study, we used the virus-induced gene silencing technology to silence the nptII gene of cottonsextracted Bt cotton and analyzed the feasibility of posttranscriptional silencing of the marker gene. The nptⅡRNAi vector was constructed by using the tobacco streak virus vector as the backbone, and cotton seedlings were infected with Agrobacterium tumefaciens. The cotton plants were infected with npt Ⅱ, and then kanamycin was applied for detection. The results showed that the nptⅡdeficient plant leaves showed macular symptoms. RT-PCR and q RT-PCR showed that the nptⅡgene transcription in leaves, roots and stems was inhibited by 99.25%, 99.05% and 98.65%, respectively, and the transcriptional inhibition in the late stage of silencing reached 98 %. The results of this study provide a quick and effective coping method and new ideas for solving the unexpected spread and unpredictable biosafety events of releasing GMOs in the environment.