【摘要】 <正>定点整合是一种指导目的基因以精确单拷贝进入基因组预定位点的转基因技术,用于解决常规转基因技术带来的系列问题^[1-2]。在拟南芥[Arabidopsis thaliana（Linn.）Heynh.]中,已通过根外植体法转化实施了基因的定点整合^[3],但这一过程存在杂菌污染、分化困难和生长周期长等问题,且定点整合效率极低[3],无法进行推广应用。花器喷雾法是一种不依赖于组织或细胞培养而直接在活体植株中进行转基因的方更多还原

【Abstract】 To improve site-specific integration technology system,site-specific integration of Rps2 target gene in Arabidopsis thaliana( Linn.) Heynh. was carried out based on Cre/lox system by floral spraying method. The results show that 1 495site-specific integration candidate plants are obtained by this method with a site-specific integration efficiency of about 0. 076%. After PCR and histochemical staining experiment verification,the positive plants of precise integration account for 86. 04%,in which,63. 34% positive plants are single copy transformed plants. The results of quantitative real-time PCR( qRT-PCR) and hypersensitive reaction( HR) show that the site-specific integrated Rps2 gene can be transcribed and expressed normally. It is suggested that this system can greatly improve the stability and efficiency of site-specific integration genetic transformation system in plants.更多还原