2019年10月17日 星期四
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 sitespecific integration of Rps2 gene in Arabidopsis thaliana based on Cre/lox system by floral spraying method
2017年 第26卷 第1期 页码[104-106]    下载全文[1.1MB]  
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.
 

关键词Cre/lox系统; Rps2基因; 定点整合; 拟南芥; 花器喷雾法
Key wordsCre/lox system; Rps2 gene; site-specific integration; Arabidopsis thaliana (Linn.) Heynh.; floral spraying method
作者李定红, 张艳梅, 周广灿, 雷照, 孙小芹, 杭悦宇
所在单位江苏省中国科学院植物研究所(南京中山植物园), 江苏 南京 210014
点击量416
基金项目江苏省科技计划项目(省属公益类科研院所能力提升)(BM2015019); 江苏省自然科学基金资助项目(BK2010476); 江苏省植物迁地保护重点实验室开放基金项目(迁201202); 江苏省中国科学院植物研究所青年基金项目(SQ201404)