莲藕多酚氧化酶互作蛋白的筛选及验证

原新博,程婷婷,惠小涵,陈章玉,王瑞红,柯卫东,郭宏波

Screening of Polyphenol Oxidase Interaction Proteins from Nelumbo nucifera and Their Verification

YUAN XinBo,CHENG TingTing,XI XiaoHan,CHEN ZhangYu,WANG RuiHong,KE WeiDong,GUO HongBo

表1 PCR 引物列表

Table 1 PCR primer list

引物名称Name 序列Sequence 用途Use APX-EcoR I-F ggaggccagtgaattcATGGCTTGTCTGGGCGGTGC 酵母双杂交 Yeast two hybrid APX-Xho I-R tcatctgcagctcgagTCATGTTGAGAAACCCTCAGGAGG NnCAT1-EcoR I-F ggaggccagtgaattcATGGATCCTTACAAGTATCGCCC NnCAT1-Xho I-R tcatctgcagctcgagTCACATGCTCGGCTTCAC cytCuZnSOD-EcoR I-F ggaggccagtgaattcATGGTGAAGGCTGTTGCGG cytCuZnSOD-Xho I-R tcatctgcagctcgagTTAACCCTGCAAGCCAATAATACC PER1-EcoR I-F ggaggccagtgaattcATGCCTGGACTGACGATCGG PER1-Xho I-R tcatctgcagctcgagTCAGACGTTGGTAAAACGGAG POD3L-EcoR I-F ggaggccagtgaattcATGAGGACTACTTATCTTCCGTT POD3L-Xho I-R tcatctgcagctcgagTTAAGGGTTCACAACCGCACACT POD43L-EcoR I-F ggaggccagtgaattcATGGCTCTGGTTTTTGCTC POD43L-Xho I-R tcatctgcagctcgagTCAATTGAAAGACCTGCAGGAC PODP7L-EcoR I-F ggaggccagtgaattcATGGCCTCCATCATCACCC PODP7L-Xho I-R tcatctgcagctcgagTCAGTTCACTCTCCTGCAATTC NnCAT116-398-EcoR I-F ggaggccagtgaattcATGTTCTGGAGCACAAATTCTGG NnCAT116-398-Xho I-R tcatctgcagctcgagTCAATCATACCTTGAAGGAAAGTAA NnCAT116-493-EcoR I-F ggaggccagtgaattcATGTTCTGGAGCACAAATTCTGG NnCAT116-493-Xho I-R tcatctgcagctcgagTCACATGCTCGGCTTCAC NnPPO1175-380-Nco I-F ctgcatatggccatgATGCCACGTAACTTCACGCA NnPPO1175-380-BamH I-R gcaggtcgacggatcTCATATTGTCCACATCCGGTCGAC NnPPO1381-468-NcoI-F ctgcatatggccatgATGTGGAAAAAGCTGGGAGG NnPPO1381-468-BamH I-R gcaggtcgacggatcTCACTCACTGACGGTTGGC NnPPO1469-597-Nco I-F ctgcatatggccatgATGTTCCCCAAAGAACTTGATGCG NnPPO1469-597-BamH I-R gcaggtcgacggatcTCACGAAGCGAACACTATCTTG NnCAT1-BD-Nco I-F ctgcatatggccatgATGGATCCTTACAAGTATCGCCC 自激活验证 Self-activated tast NnCAT1-BD-BamH I-R gcaggtcgacggatcTCACATGCTCGGCTTCAC NnPPO1-BiFC-N-BamH I-F cccaggcctactagtggatccATGGCGTCGCTGTCTCCC 双分子荧光标记 BiFC NnPPO1-BiFC-N-Xho I-R cccgggagcggtaccctcgagTCACGAAGCGAACACTATCTTGA NnCAT1-BiFC-C-BamH I-F tggcgcgccactagtggatccATGGATCCTTACAAGTATCGCCC NnCAT1-BiFC-C-Xho I-R cccgggagcggtaccctcgagCATGCTCGGCTTCACATTGA NnCAT1-GFP-BamH I-F ggactctagaggatccATGGATCCTTACAAGTATCGCCC 亚细胞定位 Subcellulcar locatization NnCAT1-GFP-Kpn I-R cccttgctcaccatggtaccCATGCTCGGCTTCACATTGAGAC β-actin-F GCCATCCAGGCCGTTCTCTC qRT-PCR β-actin-R GGGACAGTGTGGCTGACACC NnPPO1-qPCR-F CAAACTCCGCGATGCCAAGC NnPPO1-qPCR-R CGTCGAGCCATTGGACACCA NnCAT1-qPCR-F TTTGCCCTGGCGTTGTGGTC NnCAT1-qPCR-R GGTGAGCACACTTGGGAGCA