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基于拟南芥RAV类转录因子的序列,通过电子克隆的方法获得甘蓝型油菜的BnaRAV-2基因序列,再利用RT-PCR方法从甘蓝型油菜沪油15中扩增出编码RAV类转录因子基因BnaRAV-2-HY15,并进行了序列和进化分析。结果显示,该转录因子基因长1119 bp,编码372个氨基酸,含有相对保守的AP2结合域和B3结构域,具有典型的植物RAV类转录因子的结构特征。BnaRAV-2-HY15和AtRAV2具有相似的三维结构。采用半定量RT-PCR方法对基因表达水平进行分析,发现BnaRAV-2-HY15受到PEG、高盐和低温的诱导表达。BnaRAV-2-HY15基因在沪油15盛花和籽粒发育时期的根、茎、叶、花、蕾和荚中均检测不到明显的表达。
Based on the sequence of Arabidopsis RAV transcription factors, the BnaRAV-2 gene sequence of Brassica napus was obtained by electronic cloning method. The gene encoding RAV transcription factor was amplified from Brassica napus Huyou 15 by RT-PCR BnaRAV-2-HY15, and sequenced and analyzed for evolution. The results showed that the gene was 1119 bp in length and encoded 372 amino acids. The gene contained a relatively conserved AP2-binding domain and a B3 domain, with the structural characteristics of a typical plant RAV transcription factor. BnaRAV-2-HY15 and AtRAV2 have a similar three-dimensional structure. Semi-quantitative RT-PCR method was used to analyze the gene expression level, and BnaRAV-2-HY15 was induced by PEG, high salt and low temperature. The expression of BnaRAV-2-HY15 gene was not detected in roots, stems, leaves, flowers, buds and pods during the full bloom and grain development of Huyou 15.