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目的:运用电子克隆的方法获得甘蓝中AP2/ERF转录因子基因。方法:以拟南芥的AP2/ERF转录因子作为探针,对甘蓝的EST数据库进行搜索,应用相关软件进行序列拼接组装和延长。结果:克隆出甘蓝两个AP2/ERF-B3亚族转录因子Bo AP2/ERF1和Bo AP2/ERF2。通过生物信息学方法,对两条序列编码蛋白质的氨基酸组成,亲水性/疏水性、理化性质、亚细胞定位、高级结构和空间构型等方面进行了预测和分析。结论:Bo AP2/ERF1和Bo AP2/ERF2基因由371和352个氨基酸组成、相对分子质量为40.85k Da和39.44k Da的蛋白质,定位于细胞核。序列分析结果显示,该蛋白可能具有信号转导和胁迫响应应答等功能。
Objective: To obtain the AP2 / ERF transcription factor gene in cabbage by electronic cloning method. Methods: The AP2 / ERF transcription factor of Arabidopsis thaliana was used as a probe to search the EST database of Brassica oleracea L., and the related software was used to assemble and extend the EST database. Results: Two AP2 / ERF-B3 subfamily transcription factors, Bo AP2 / ERF1 and Bo AP2 / ERF2, were cloned from Brassica. Bioinformatics methods were used to predict and analyze the amino acid composition, hydrophilicity / hydrophobicity, physico-chemical properties, subcellular localization, high-order structure and spatial configuration of the two encoded proteins. CONCLUSION: The Bo AP2 / ERF1 and Bo AP2 / ERF2 genes are composed of 371 and 352 amino acids with relative molecular weights of 40.85 kDa and 39.44 kDa and localized in the nucleus. Sequence analysis showed that the protein may have functions of signal transduction and stress response.