甘蓝型油菜BnaLCR23基因CRISPR-Cas9表达载体的构建及遗传转化

发布时间：2018-03-04 05:05

本文选题：甘蓝型油菜　切入点：BnaLCR　出处：《分子植物育种》2017年08期 　论文类型：期刊论文

【摘要】：BnaLCR23基因是甘蓝型油菜"湘油15号"种子不同发育时期抑制差减杂交文库中的一个功能未知基因,分析表明其功能可能与种子发育和油脂代谢相关,为了明确该基因的功能,本研究基于CRISPR-Cas9基因编辑系统构建了该基因的敲除质粒。利用根癌农杆菌介导的油菜下胚轴遗传转化方法将敲除质粒转入甘蓝型油菜中,经潮霉素抗性筛选以及抗性苗基因组DNA的PCR检测,获得转基因油菜。转基因油菜材料的获得为深入研究BnaLCR23基因在甘蓝型油菜种子发育和储存物的积累过程中的功能提供了帮助。
[Abstract]:BnaLCR23 gene is an unknown gene in the suppression subtractive hybridization library of "Xiangyou 15" seeds in different developmental stages of Brassica napus. The analysis shows that its function may be related to seed development and lipid metabolism. In this study, the knockout plasmid was constructed based on CRISPR-Cas9 gene editing system. The knockout plasmid was transferred into Brassica napus by Agrobacterium tumefaciens mediated Hypocotyl genetic transformation. Transgenic rapeseed was obtained by hygromycin resistance screening and PCR detection of genomic DNA of resistant seedlings. The obtained transgenic rapeseed materials provided help for further study on the function of BnaLCR23 gene in the seed development and storage accumulation of Brassica napus L.
【作者单位】：湖南农业大学生物科学技术学院;湖南农业大学农学院;
【基金】：国家自然基金面上项目(31571707) 国家重点研发计划项目子课题(2016YFD0100202-8)共同资助
【分类号】：Q943.2;S565.4

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