采用花器喷雾法并基于Cre/lox系统定点整合拟南芥Rps2基因的技术体系研究

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 1495 site?specific integration candidate plants are obtained by this method with a site?specific integration efficiency of about 0076%. After PCR and histochemical staining experiment verification, the positive plants of precise integration account for 8604%, in which, 6334% 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.