利用CRISPR/Cas9技术研究玉米ZmFKF1在开花过程中的作用

杨敏,胥华伟,王翠玲,杨护,魏岳荣

Using CRISPR/Cas9-mediated Targeted Mutagenesis of ZmFKF1 Delayed Flowering Time in Maize

YANG Min,XU HuaWei,WANG CuiLing,YANG Hu,WEI YueRong

表1 试验所用的克隆、载体构建和鉴定引物

Table 1 Cloning and identification primers used in this study

引物名称 Primer name 引物序列 Primer sequence (5′-3′) 用途 Usage ZmFKF1-F CACCATGTTTGACGACGCGGGGGC ZmFKF1克隆 Cloning of ZmFKF1 gene ZmFKF1-R GACTTTGCCATGCGAATCTACAATGC FKF1-U6b-F GTTGAGGTGGACGCGGAGCCGGGA 载体构建及鉴定 Vector construction and verification FKF1-U6b-R AAACTCCCGGCTCCGCGTCCACCT U-F CTCCGTTTTACCTGTGGAATCG gR-R CGGAGGAAAATTCCATCCAC Pps-R TTCAGAggtctcTaccgACTAGTCACGCGTATGGAATCGGCAGCAAA Pgs-L AGCGTGggtctcGctcgACGCGTATCCATCCACTCCAAGC SP-L1 GCGGTGTCATCTATGTTACTAG SP-R TGCAATAACTTCGTATAGGCT Cas9-F CTGACGCTAACCTCGACAAG 鉴定Cas9 PCR detection for Cas9 Cas9-R CCGATCTAGTAACATAGATGACACC ZmCri-F CCCTACCTGGCAGCAGAATC 靶点检测及测序引物 Target detection and sequencing primer ZmCri-R AGCAGCCCAAATCTGTCAGTC ZmUbi-F GTCTCCGTGGTGGTCAGTAAGT qRT-PCR检测引物 Primers used for qRT-PCR assay ZmUbi-R GACACGAACAGCAGATACTTTGAC ZmGI-F ACTTTCCCAACCAACGC ZmGI-R GGGCAGCTACAACAATTTCA conz1-F ATCGTCAATGGAGGTGGGTAT conz1-R GACCCGACGAGAACAGGC ZmZCN8-F ACTTGCACTGGATGGTGACAGATA ZmZCN8-R CGGCCTCTCATAAAATATTAGCTCTT