基于CRISPR/Cas9技术的水稻温敏不育基因tms5突变体的构建

黄忠明,周延彪,唐晓丹,赵新辉,周在为,符星学,王凯,史江伟,李艳锋,符辰建,杨远柱

Construction of tms5 Mutants in Rice Based on CRISPR/Cas9 Technology

Zhong-Ming HUANG,Yan-Biao ZHOU,Xiao-Dan TANG,Xin-Hui ZHAO,Zai-Wei ZHOU,Xing-Xue FU,Kai WANG,Jiang-Wei SHI,Yan-Feng LI,Chen-Jian FU,Yuan-Zhu YANG

图3 酶切和PCR鉴定pCAMBIA1301-Cas9-TMS5-gRNA载体 A: Kpn I和Hind III酶切鉴定pCAMBIA1301-Cas9-TMS5-gRNA载体。箭头所示为CRISPR/Cas9系统元件的片段; B: 利用GUS-JC-F和GUS-JC-R引物PCR鉴定农杆菌中的pCAMBIA1301-Cas9-TMS5-gRNA载体。M: DL2000; 1: 18T-Cas9-TMS5-gRNA; 2: pCAMBIA1301; 3: pCAMBIA1301-Cas9-TMS5-gRNA。

Fig. 3 Identification of the pCAMBIA1301-Cas9-TMS5-gRNA plasmid by restriction enzyme digestion and PCR A: identification of the pCAMBIA1301-Cas9-TMS5-gRNA plasmid digested with Kpn I and Hind III; B: identification of the pCAMBIA1301-Cas9-TMS5-gRNA plasmid in Agrobacterium tumefaciens by PCR with primers of GUS-JC-F and GUS-JC-R. M: DL2000; 1: 18T-Cas9-TMS5-gRNA; 2: pCAMBIA1301; 3: pCAMBIA1301-Cas9-TMS5-gRNA.