甘蓝型油菜BnMAPK1的原核表达、亚细胞定位及酵母双杂交文库筛选

王珍, 姚梦楠, 张晓莉, 曲存民, 卢坤, 李加纳, 梁颖

Prokaryotic expression, subcellular localization and yeast two-hybrid library screening of BnMAPK1 in B. napus

Zhen WANG, Meng-Nan YAO, Xiao-Li ZHANG, Cun-Min QU, Kun LU, Jia-Na LI, Ying LIANG

表1 所用PCR引物

Table 1 PCR primers used in this study

引物名称 Primer name 引物序列 Primer sequences (5°-3°) 用途 Usage P-BnMAPK1 F: GGATCCATGGCGACACCAGTTGATC R: GTCGACTAGCAGCTCAGATGTTGA BnMAPK1的原核表达载体构建 Construction of prokaryotic expression vector of BnMAPK1 T7 F: TAATACGACTCACTATAGGG R: TGCTAGTTATTGCTCAGCGG 原核表达重组载体的检测 Detection of prokaryotic expression recombinant vectors SC-BnMAPK1 F: ACGAATTCATGGCGACACCAGTTGATC R: ACGGATCCTAGCAGCTCAGATGTTGA BnMAPK1的亚细胞定位表达载体构建 Construction of subcellular localization vector of BnMAPK1 P389 F: ACATGGTCCTGCTGGAGTTC R: ATTGCCAAATGTTTGAACGA 亚细胞定位重组载体的检测 Detection of subcellular localization recombinant vectors eGFP F: ATGGTGAGCAAGGGCGAGGAG R: GGACTTGTACAGCTCGTCCATGCC 亚细胞定位重组载体的检测 Detection of subcellular localization recombinant vectors Y2H-BnMAPK1 F: CGGAATTCATGGCGACACCAGTTGATC R: CGGGATCCTCATAGCAGCTCAGATGT BnMAPK1的酵母双杂交载体构建 Construction of Yeast Two-Hybrid vector of BnMAPK1 BD F: TAATACGACTCACTATAGGG R: TTTTCGTTTTAAAACCTAAGAGTC Bait载体的检测 Detection of Bait vectors AD F: TAATACGACTCACTATAGGG R: AGATGGTGCACGATGCACAG 文库Prey载体的检测 Detection of Prey vectors in library