大豆生物钟基因GmLNK1/2、GmRVE4/8和GmTOC1 CRISPR/Cas9组织表达分析及敲除靶点的鉴定

甘卓然,石文茜,黎永力,侯智红,李海洋,程群,董利东,刘宝辉,芦思佳

Identification of CRISPR/Cas9 knockout targets and tissue expression analysis of circadian clock genes GmLNK1/2, GmRVE4/8, and GmTOC1 in soybean

Zhuo-Ran GAN,Wen-Qian SHI,Yong-Li LI,Zhi-Hong HOU,Hai-Yang LI,Qun CHENG,Li-Dong DONG,Bao-Hui LIU,Si-Jia LU

表1 基因靶点引物列表

Table 1 List of gene target primers

靶点引物 Target primer 引物序列 Primer sequence (5°-3°) 靶点对应基因 Target of gene LNK1T1F gtcaGGAGACAAGTGTGTGGTGG GmLNK1a, GmLNK1b LNK1T1R aaacCCACCACACACTTGTCTCC GmLNK1a, GmLNK1b LNK1T2F gtcaAGAGGAGTTCTGCTGGCTC GmLNK1a, GmLNK1b LNK1T2R aaacGAGCCAGCAGAACTCCTCT GmLNK1a, GmLNK1b LNK1T3F attgCTTGGGAGACAAGTGTGTGG GmLNK1c, GmLNK1d LNK1T3R aaacCCACACACTTGTCTCCCAAG GmLNK1c, GmLNK1d LNK1T4F attgAGACTTTGAAGATGTTGAC GmLNK1c, GmLNK1d LNK1T4R aaacGTCAACATCTTCAAAGTCT GmLNK1c, GmLNK1d LNK2T1F gtcaACATAATATGGGGTGAAGG GmLNK2a, GmLNK2b LNK2T1R aaacCCTTCACCCCATATTATGT GmLNK2a, GmLNK2b LNK2T2F gtcaAAACTGATCAGGGTTCCCT GmLNK2c, GmLNK2d LNK2T2R aaacAGGGAACCCTGATCAGTTT GmLNK2c, GmLNK2d LNK2T3F attgTTTGATTGGAACGACGAAG GmLNK2a, 2b, 2c, 2d LNK2T3R aaacCTTCGTCGTTCCAATCAAA GmLNK2a, 2b, 2c, 2d LNK2T4F attgTCATATTGTGCCTTATCCGG GmLNK2c, GmLNK2d LNK2T4R aaacCCGGATAAGGCACAATATGA GmLNK2c, GmLNK2d RVE48T1F gtcaCTTCCCTGCTGATGAATGC GmRVE4/8b, GmRVE4/8c RVE48T1R aaacGCATTCATCAGCAGGGAAG GmRVE4/8b, GmRVE4/8c RVE48T2F gtcaTCATCCCATGTGACATACCC GmRVE4/8a, GmRVE4/8d RVE48T2R aaacGGGTATGTCACATGGGATGA GmRVE4/8a, GmRVE4/8d RVE48T3F attgCAGCTTTGCGCTTTGGACG GmRVE4/8b, GmRVE4/8c RVE48T3R aaacCGTCCAAAGCGCAAAGCTG GmRVE4/8b, GmRVE4/8c RVE48T4F attgAAGCTTTGCGCTTAGGCCG GmRVE4/8a, GmRVE4/8d RVE48T4R aaacCGGCCTAAGCGCAAAGCTT GmRVE4/8a, GmRVE4/8d TOC1T1F gtcaCGATTCCAAGAGTTCTCAAG GmTOC1a, GmTOC1b TOC1T1R aaacCTTGAGAACTCTTGGAATCG GmTOC1a, GmTOC1b TOC1T2F gtcaGTGATGTCCGCACAAGATG GmTOC1a, GmTOC1b TOC1T2R aaacCATCTTGTGCGGACATCAC GmTOC1a, GmTOC1b TOC1T3F attgGTGGGAATAATAGTAAGAG GmTOC1c, GmTOC1d TOC1T3R aaacCTCTTACTATTATTCCCAC GmTOC1c, GmTOC1d TOC1T4F attgTTGTAAAGTGCTTGAGGCT GmTOC1c, GmTOC1d TOC1T4R aaacAGCCTCAAGCACTTTACAA GmTOC1c, GmTOC1d