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
Technology system research on sitespecific integration of Rps2 gene in Arabidopsis thaliana based on Cre/lox system by floral spraying method
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 1 495 site-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.
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