论文部分内容阅读
CRISPR/Cas9 (Clustered regulatory interspaced short palindromic repeat/Cas9)基因编辑技术在生命科学领域掀起了一场重大的技术革命,它比锌指核酸酶(ZFNs)和转录激活因子效应物核酸酶(TALENs)技术更易于操作,而且更高效.CRISPR/Cas9系统已经被广泛应用到植物科学研究领域中.本实验选取拟南芥MS2为目的基因,构建植物基因编辑系统的表达载体,并通过农杆菌介导的方法转化拟南芥,从而利用CRISPR/Cas9系统靶向敲除拟南芥MS2基因.对转基因后代的MS2测序结果分析表明,在获得的12个阳性转化植株中,发现了5个阳性植株存在基因序列上的新碱基插入,由此形成CRISPR/Cas9介导的拟南芥MS2突变体.这一工作为后续分析其他物种中MS2同源基因的功能研究提供参考依据.“,”The clustered regulatory interspaced short palindromic repeat/Cas9 (CRISPR/Cas9) genome editing technology launches a great technological revolution in the field of life sciences. Compared with zinc finger nucleases (ZFNs) and transcription activator-like effector nucleases (TALENs) technologies, the CRISPR/Cas9 technology is easier to operate and more efficient, CRISPR/Cas9 system has been widely applied in the field of plant science. MS2 gene of Arabidopsis thaliana was selected as the target gene in this study, and the expression vector of plant gene editing system was constructed. Then, the Arabidopsis thaliana was transferred by Agrobacterium-mediated method, and the MS2 gene of Arabidopsis thaliana was knocked out by CRISPR/Cas9 system. The results of MS2 sequencing of transgenic progeny showed that in the 12 positive transgenic plants, 5 positive plants were found to have new bases inserted in the gene sequence, thus forming a CRISPR/Cas9 mediated Arabidopsis MS2 mutant. This study provided a basis for subsequent functional analysis of MS2 homologous genes in other species.