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RNA干扰(RNAi)技术是研究基因功能的一种常用方法.为快速检测RNAi载体的沉默效果,通过构建木薯eIF4E3基因的过量表达载体pAI-Ca4E3以及靶标木薯eIF4E3基因的RNAi表达载体p1300-Ca4E3,利用农杆菌注射法将表达载体在本生烟叶片中进行瞬时表达.半定量RT-PCR检测结果显示单独注射过量表达载体pAI-Ca4E3或与表达载体pCAMBIA 13 00共同注射的样品,农杆菌注射后第4天、第5天和第6天都能检测到木薯eIF4E3基因的高效表达,并且二者木薯eIF4E3基因表达量基本相同,说明木薯eIF4E3基因能在本生烟中高效瞬时表达,但是过量表达载体pAI-Ca4E3与干扰载体p1300-Ca4E3共同注射的样品,农杆菌注射后第4天、第5天还是第6天后都几乎检测不到木薯eIF4E3基因的表达,说明过量表达载体表达的木薯eIF4E3基因己被干扰载体有效沉默.研究结果表明,己建立同时表达靶标基因和干扰靶标基因的干扰载体的本生烟瞬时表达系统,并结合半定量RT-PCR进行RNAi载体沉默效果检测的方法.该方法在农杆菌注射4d后就能有效的检测RNAi载体的沉默效果,具有快捷、操作简便等特点,为RNAi技术应用于基因功能研究提供简单、快捷、有效的证据.“,”RNA interference (RNAi) technique has been used in gene functional research frequently.In order to identify the silencing effects of RNAi vector,expression vector pAI-Ca4E3 which over-expressing cassava eIF4E3 gene and RNAi expressing vector p1300-Ca4E3 which harborring hairpin structure targeting cassava eIF4E3 gene were constructed and transient expressed in Nicotiana banthemiana through Agrobacteria mediating infiltration.The semi-quantitative RT-PCR results showed that the expression level of samples infiltrated with construct pAI-Ca4E3 alone and coinfiltrated with construct pCAMBIA1300 were similar,no matter 4 d post infiltration (dpi),5 dpi or 6 dpi,which indicated the cassava eIF4E3 gene could be transient expressed efficiently in N.banthemiana.However,the expression of cassava eIF4E3 gene almost undetectable in the samplesthat co-infiltrated with construct pAI-Ca4E3 and RNAi expressing vector p 1300-Ca4E3,no matter 4 d post infiltration (dpi),5 dpi or 6 dpi,which showed the cassava eIF4E3 gene has been silenced efficiently by RNAi expressing vector p1300-Ca4E3.All together the results showed that we established a fast and efficient method to identify the efficiency of RNAi construct by employing transient expression system of N.Banthemiana combining with semi-quantitative RT-PCR.The method can effectively detect the silencing effects of RNAi vector afterinjection agrobacterium for 4 d,the detection method could provide a simple,quick and effective evidence for the application of RNAi technology in gene function research.