基于CRISPR/Cas9系统构建拟南芥EXPA多基因编辑表达载体  

The Construction of Multi-gene Editing Expression Vector of ArabidopsisEXPA by CRISPR/Cas9 System

作　　者：廖嘉明 包钰韬 陈媛 李布野 李华强 欧阳昆唏 陈晓阳[1,2] Liao Jiaming;Bao Yutao;Chen Yuan;Li Buye;Li Huaqiang;Ouyang Kunxi;Chen Xiaoyang(Guangdong Key Laboratory for Innovative Development and Utilization of Forest plant Germplasm,College of Forestry and Landscape Architecture of South China Agriculture University,Guangzhou,510642;State Key Laboratory for Conservation and Utilization of Subtropical Agro-bioresources,South China Agricultural University,Guangzhou,510642;China Forestry Group Leizhou Forestry Bereau Co.,Ltd.,Zhanjiang,524348)

机构地区：[1]华南农业大学林学与风景园林学院,广东省森林植物种质创新与利用重点实验室,广州510642 [2]华南农业大学亚热带农业生物资源保护与利用国家重点实验室,广州510642 [3]中林集团雷州林业局有限公司,湛江524348

出　　处：《分子植物育种》2020年第17期5748-5754,共7页Molecular Plant Breeding

基　　金：国家自然科学基金(31600525);广东省科技计划项目(2017B020201008);中央财政林业科技推广示范项目((2018)GDTK-08)共同资助。

摘　　要：CRISPR/Cas9系统是一种由sg RNA及Cas9核酸酶组成的适应性免疫系统,已被成功运用于多种植物基因组编辑。扩展蛋白(expansin, EXP)是一类可以使细胞壁伸展和松弛的细胞壁蛋白,对植物的各个组织生长发育都有着重要的作用,由EXPA、EXPB、EXLA和EXLB 4个不同的基因家族组成。本研究以拟南芥的EXPA基因为例,通过CRISPR/Cas9系统构建拟南芥EXPA多基因编辑的表达载体,并利用农杆菌介导CRISPR/Cas9表达载体的方法转化拟南芥,以创制拟南芥扩展蛋白多基因突变体,为后续的研究提供突变体材料。The CRISPR/Cas9 system is an adaptive immune system consisting of sgRNA and Cas9 nuclease.It has been successfully applied to a variety of plant genome editing.Expansin(EXP)is a kind of cell wall protein that function in cell wall extension and relaxation.It plays important roles in the growth and development of various tissues of plants.It consists of four different gene families:EXPA,EXPB,EXLA and EXLB.Based on the EXPA genes of Arabidopsis thaliana,the research constructed the multi-gene editing expression vector of EXPA in Arabidopsis thaliana by CRISPR/Cas9 system.Moreover,Agrobacterium-mediated CRISPR/Cas9 expression vector was used to transform Arabidopsis thaliana,in order to create multi-gene mutant of Arabidopsis thaliana and provide mutant materials for subsequent research.

关 键 词：拟南芥杂交 CRISPR/Cas9 基因编辑 扩展蛋白 

分 类 号：Q943.2[生物学—植物学]