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 words | Cre/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 |
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| 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) |