拟南芥过氧化氢酶敲除载体构建

Establishment of catalase knockout vector system in Arabidopsis thaliana

过氧化氢酶(CAT)是一种重要的H_2O_2清除酶,广泛地存在于有氧生物细胞体内,并且对于维持体内氧化水平至关重要。因此,获得完全敲除CAT遗传突变体无疑对于研究H_2O_2调控植物生长和发育提供重要的材料。目前已知拟南芥中有3个CAT基因(CAT1、CAT2、CAT3),其中CAT1和CAT3两个基因之间的连锁遗传,很难获得使CAT完全缺失突变体(cat1 cat2 cat3)。文章利用CRISPR/Cas9技术构建多靶点双元载体以期同时敲除CAT1、CAT3基因,并转入cat2突变体中,以期获得cat1 cat2 cat3三突变体。CAT完全缺失突变体的获得为研究过氧化氢酶功能及H_2O_2信号转导机制提供必要的遗传材料。

Catalase（CAT）,an important H2O2 scavenger,is found in aerobic living cells and is essential for the maintenance of oxidative levels in vivo.Therefore,the complete knockout of CAT genetic mutants will undoubtedly provide important materials for studying the regulation of H2O2 on plant growth an development.It is known that there are three CAT genes（CAT1,CAT2,CAT3）in Arabidopsis thaliana.It is difficult to obtain the complete CAT deletion mutant（cat1 cat2 cat3）due to the linkage between CAT1 and CAT3 genes.In this paper,a multi-target binary vector was constructed by using CRISPR/Cas9 technology to knock out CAT1 and CAT3 genes simultaneously,and then transferred into cat2 deletion mutant to obtain cat1 cat2 cat3 deletion mutant.The complete deletion mutant of CAT will provide the necessary genetic material for the study of CAT function and H2O2 signal transduction mechanism.